Azure Sapphire FL Biomolecular Imager with lid open

Sapphire FL Biomolecular Imager

IS4000

Designed for flexible choice in detection chemistry and samples, the Sapphire FL brings precise quantitation of nucleic acids and proteins

SCA_NomineeBadge_StrokeGold24

Unmatched Application Versatility and Performance

laser-based imaging

24-bit Dynamic Range

 

Extended dynamic range allows imaging of both bright and weak bands without experiencing saturation.

5 micron resolution icon

5 Micron Resolution


From Western blots and gels, to microscope slides and live animal imaging, adjustable 5-1000 µm resolution ensures unparalleled flexibility.

quantity accuracy

Custom Design Lasers and Filters

No limits – easily swap lasers and filters to achieve the perfect system to image your sample

Adjustable focus Sapphire FL

Adjustable focus for sample thickness

 

Whether imaging Western blots, gels, 96-well plates, or tissue slides, the adjustable Z-plane ensures optimal focus for your sample.

Key Features

Flexibility with uncompromising performance – From in vitro molecular assays to in vivo imaging

The Sapphire FL is the second generation Sapphire and ultimate biomolecular imager for FLEXIBILITY. With customizable and user-changeable laser and filter modules, it easily adapts to a lab’s changing needs and advancing research. The Sapphire FL offers customizable and user-changeable optical modules, 5–1000 μm resolution scans, a Z-plane range from -1.0 to +6 mm, 5 anesthesia ports for imaging living animals, chemiluminescence detection through a Chemiluminescence Module, and much more.

Unlimited Laser and Filter Wavelength combinations

Scientist changing optical modules on the new Azure Sapphire FL
Pick the modules that support your research. Changing the optical modules on the new Sapphire FL is simple and easy. The unique mechanism makes selecting lasers to match your dyes finally possible. Easily swap lasers, filters, and/or entire optical modules in under two minutes to suit the needs of your experiment.

The Sapphire FL is the only biomolecular scanner in its class that reaches into the UV spectrum, allowing multiplexing of UV dyes with dyes in the NIR. Using the 375 Standard Optical Module, it is able to image common DNA stains, such as Dapi and Hoechst.

It features a unique, patent-pending design of interchangeable and customizable laser and filter modules, enabling a virtually infinite number of spectral combinations. A broad range of excitation and emission wavelengths, as well as phosphor imaging, are supported.

This versatility permits research-driven, rather than instrument-dictated, experimental design. With the Sapphire FL, the user-exchangeable optical modules (consisting of one laser and one emission filter) ensure there is no need to compromise.

Sapphire FL Applications
The Sapphire FL boasts the widest wavelength range of any biomolecular imaging system with the support of wavelengths between 375 – 900 nm, allowing it to support imaging of Western blots, gels, phosphor screens, tissues, slides, arrays and much more.

Application Flexibility

Sapphire FL Applications
The Sapphire FL boasts the widest wavelength range of any biomolecular imaging system with the support of wavelengths between 375 – 900 nm, allowing it to support imaging of Western blots, gels, phosphor screens, tissues, slides, arrays and much more.

The flexibility of the Sapphire FL guarantees researchers the freedom to confidently choose the best dyes and chemistry for any experiment. Custom optical modules can excite any dye at its peak, resulting in the highest possible signal for any fluorophore.

In addition to fluorescence, the Sapphire FL also excels at imaging phosphor screens and chemiluminescence using the optional Chemiluminescence Module.

Sensitive Fluorescent Detection

Bovine serum albumin (BSA) conjugated to AzureSpectra dyes, separated by SDS-PAGE, and transferred to membranes.
Bovine serum albumin (BSA) conjugated to AzureSpectra dyes, separated by SDS-PAGE, and transferred to membranes. Western blots were imaged at 50 μm on the Sapphire FL. Loaded amounts of dye-conjugated-BSA are given.

High sensitivity allows femtogram detection of proteins labeled with common fluorescent dyes.

No other system on the market matches the Sapphire FL’s coverage across the fluorescent spectrum. Extend your reach into the UV spectrum with our 375 Standard Optical Module.

Bovine serum albumin (BSA) conjugated to AzureSpectra dyes, separated by SDS-PAGE, and transferred to membranes.
Bovine serum albumin (BSA) conjugated to AzureSpectra dyes, separated by SDS-PAGE, and transferred to membranes. Western blots were imaged at 50 μm on the Sapphire FL. Loaded amounts of dye-conjugated-BSA are given.
American Radiolabel Chemicals Carbon-14 standard exposed to Storage Phosphor Screen for three hours, then imaged with the Azure Sapphire Biomolecular Imager
American Radiolabel Chemicals Carbon-14 standard exposed to Storage Phosphor Screen for three hours, then imaged with the Sapphire
Sample: C autoradiographic standard; Imaging; Phosphor; LOD: 0.036 µCi/g; DR: 5.4 orders of magnitude; Linearity: R²=0.99
Plot showing linearity and linear dynamic range achieved when a carbon-14 standard was imaged using the Sapphire

Total Flexibility with Phosphor Imaging

American Radiolabel Chemicals Carbon-14 standard exposed to Storage Phosphor Screen for three hours, then imaged with the Azure Sapphire Biomolecular Imager
American Radiolabel Chemicals Carbon-14 standard exposed to Storage Phosphor Screen for three hours, then imaged with the Sapphire Biomolecular Imager
Sample: C autoradiographic standard; Imaging; Phosphor; LOD: 0.036 µCi/g; DR: 5.4 orders of magnitude; Linearity: R²=0.99
Plot showing linearity and linear dynamic range achieved when a carbon-14 standard was imaged using the Sapphire

The Sapphire FL is a compact phosphor imager that incorporates exceptional visible fluorescent and near infra-red scanning along with true chemiluminescent and white light imaging, to make it a true lab workhorse.

It is one of only two scanning systems in its class that offers phosphor imaging, a high sensitivity assay that requires radioactivity. Sapphire FL is able to scan storage phosphor screens with exceptional dynamic range and image quality, thanks to the use of laser excitation and photon multiplier tube (PMT) detection.

In Vivo Imaging

Several attributes of the Sapphire FL imager make imaging of live animals possible. When combined with appropriate tubing and nose cones, living mice can be fluorescently imaged under anesthesia using one of five built-in anesthesia ports.

The wide imaging platform can accommodate specimens with depth (up to 4 cm) allowing mice, plants, petri dishes, and other large biological samples to be imaged.

Scanning with EDR demonstrates quantifiable linearity over the entire range of sample concentrations.
Dot blot scanned with (top image) and without (bottom image) EDR. Without EDR, the top four dots saturate and cannot be quantified. Scanning with EDR demonstrates quantifiable linearity over the entire range of sample concentrations.

Distinguish between Subtle Differences in Expression with Extended Dynamic Range (EDR)

Scanning with EDR demonstrates quantifiable linearity over the entire range of sample concentrations.
Dot blot scanned with (top image) and without (bottom image) EDR. Without EDR, the top four dots saturate and cannot be quantified. Scanning with EDR demonstrates quantifiable linearity over the entire range of sample concentrations.

Extended dynamic range, when selected, allows imaging of both bright and weak bands without experiencing saturation. This is ideal for samples that feature strong and weak expressing proteins.

EDR extends dynamic range to 24 bits of data.

A 50-micron scan of a flower was taken using an Azure Sapphire FL from 0 to 6 mm with 1 mm adjustments.
A. 50-micron scan of a flower was taken from 0 to 6 mm with 1 mm adjustments.
50-micron scan of the same flower was merged in the Sapphire FL Capture Software to view all areas of focus at once.
B. 50-micron scan of the same flower was merged in the Sapphire FL Capture Software to view all areas of focus at once.

Adjustable Focus

We understand some samples are on the glass (Western blots), while some samples are raised above the glass surface (96-well plates). Some samples also have multiple focal points within them, such as tissue slides and organs. Sapphire FL can image those too.

The Sapphire FL offers a 7mm software-controlled adjustable focal plane, so you can scan your sample at multiple depths to ensure you see all your data has to offer.

5-micron Resolution

Mouse lung tissue slide imaged on the Sapphire FL with 638 nm laser modules at 5 μm.
Mouse lung tissue slide probed for vascular endothelial (VE)-cadherin (AzureSpectra 550 nm secondary antibodies) and smooth muscle actin (SMA) (AzureSpectra 650 nm secondary antibodies). Imaged on the Sapphire FL using the 532 and 638 Standard Optical Modules (red and green, respectively) at 5 um.

 The Sapphire FL Z-plane ranges from -1 mm below to +6 mm above the glass imaging surface, allowing crystal clear imaging of samples with depth. The adjustable focal plane enables scanning of thick samples. Screen slides before microscopic analysis by imaging multiple slides at a resolution of 5 microns. 

Optimize the focal plane for your sample. Easily switch between sample types with confidence that each image will be sharply in focus. Create Z-stacks and GIFs to visualize multiple focal planes at once.

Mouse lung tissue slide imaged on the Sapphire FL with 638 nm laser modules at 5 μm.
Mouse lung tissue slide probed for vascular endothelial (VE)-cadherin (AzureSpectra 550 nm secondary antibodies) and smooth muscle actin (SMA) (AzureSpectra 650 nm secondary antibodies). Imaged on the Sapphire FL using the 532 and 638 Standard Optical Modules (red and green, respectively) at 5 um.
chemiluminescent FLASH Western Blot HeLa serial each in duplicate antitubulin antiactin with marker imaged with Azure Biosystems chemiSOLO
Western blot of serially diluted HeLa cell lysate in duplicate with marker, probed for Tubulin and Actin, detected with the Flash Western Chemiluminescence Kit. Visualized on the Sapphire FL Chemiluminescence Module.
chemiluminescent FLASH Western Blot HeLa serial each in duplicate antitubulin antiactin with marker imaged with Azure Biosystems chemiSOLO
Western blot of serially diluted HeLa cell lysate in duplicate with marker, probed for Tubulin and Actin, detected with the Flash Western Chemiluminescence Kit. Visualized on the Sapphire FL Chemiluminescence Module.

Chemiluminescent Imaging When You Need It

The Chemiluminescence Module adds high-resolution, quantitative chemiluminescence and visible imaging.

The chemi module allows you to express proteins with femtogram sensitivity and gives you the ability to capture marker images.

Trusted by Scientists Worldwide

Applications

The new Sapphire FL provides you with the versatility to do more applications with one system. Browse the list of published examples below to see what the SFL is capable of.

Fluorescence Imaging

White Light and Autofluorescence

Rescue of autoimmune hepatitis by soluble MHC class II molecules in an altered concanavalin A-induced experimental model

Katerina Bakela, Maria Georgia Dimitraki, Evangelia Skoufa, Irene Athanassakis

…Liver tissues were isolated 6 months after the ConA and the ConA + sMHCII treatment of mice and fixed using PFA 4%, under rotation, at 4°C for 24 hours. Azure Biosystems Sapphire™ Biomolecular Imager (Azure Biosystems, Dublin, CA 94568 USA) was used in order to scan the liver tissues in 10-μm resolution. This instrument combines NIR fluorescence (both long and short), RGB fluorescence, chemiluminescent, and phosphor imaging capabilities, while using four solid state lasers as excitation sources (450, 520, 660, and 780 nm). The application of the four-channel fluorescence mode at 10-μm resolution, could detect gross anatomy and morphology of liver tissues, mainly based on tissue autofluorescence…

Figure 6. Livers were isolated, fixed and scanned using a white light beam at 10-μm resolution (Ca) or four-channel fluorescence at 10-μm
resolution (Cb).
Figure 6. Livers were isolated, fixed and scanned using a white light beam at 10-μm resolution (Ca) or four-channel fluorescence at 10-μm resolution (Cb).
Immunohistochemistry

The ISL LIM-homeobox 2 transcription factor is negatively regulated by circadian adrenergic signaling to repress the expression of Aanat in pinealocytes of the rat pineal gland

Mattern KMJ, Blancas–Velázquez AS, Ngo MT, et al.

Cryostat sections of 12-μm thickness were mounted on slides and blocked in 5% normal donkey serum (Abcam). Immunohistochemistry was performed as previously described…Sections were incubated in primary antibody at 4°C overnight and in secondary antibody at room temperature for…1 h if used for whole section imaging…Images of whole sections were taken on an Azure Sapphire Biomolecular Imager (Azure Biosystems). Fluorescence signal intensity after background substraction [sic] was quantified with the Azure Spot software… “Western blot” for image acquisition, an Azure Sapphire Biomolecular Imager was used. The density of bands was quantified with the Azure Spot software.

J Pineal Res. 2023;e12905
Figure 3. Diurnal dynamics of Isl2 expression in the pineal gland of the adult rat…(C) Two representative western blot lanes of samples containing protein extracted from pineal glands were harvested at ZT10 and ZT22 to visualize the difference of ISL2 abundancy in daytime versus nighttime samples. Rat ISL2 protein (NP_065204) has a predicted molecular weight of 39.7 kDa. Housekeeping proteins ACTB and GAPDH were used as loading controls…(D) Two representative immunohistochemical stainings in brain sections from brains removed at ZT10 and ZT22 to visualize differences in ISL2 abundancy. Scale bars = 1 mm. (E) Quantification of ISL2 levels in the individual lanes of the western blot shown in (C).
Figure 3. Diurnal dynamics of Isl2 expression in the pineal gland of the adult rat…(C) Two representative western blot lanes of samples containing protein extracted from pineal glands were harvested at ZT10 and ZT22 to visualize the difference of ISL2 abundancy in daytime versus nighttime samples. Rat ISL2 protein (NP_065204) has a predicted molecular weight of 39.7 kDa. Housekeeping proteins ACTB and GAPDH were used as loading controls…(D) Two representative immunohistochemical stainings in brain sections from brains removed at ZT10 and ZT22 to visualize differences in ISL2 abundancy. Scale bars = 1 mm. (E) Quantification of ISL2 levels in the individual lanes of the western blot shown in (C).

laser scanning, ISL2 was detected using a sheep anti-hlslet-2 primary antibody and an Alexa Fluor™ 790 donkey anti-sheep secondary antibody. For Western blotting, ISL2 was detected with a sheep-anti-human Islet-2 antibody and a donkey-anti-sheep Alexa Fluor™ 680 secondary antibody. ACTB was detected with a rabbit-anti-beta actin antibody and a donkey-anti-rabbit Alexa Fluor™ 790 secondary antibody.

Parallel imaging of coagulation pathway proteases activated protein C, thrombin, and factor Xa in human plasma

Sylwia Modrzycka, Sonia Kołt, Stéphanie G. I. Polderdijk, Ty E. Adams, Stanisław Potoczek, James A. Huntington, Paulina Kasperkiewicz and Marcin Drąg

 …For the simultaneous coagulation factor labeling, human plasma was incubated with 5 μM of each fluorescently labeled probe…and then 5 μL of each sample was run onto a 10% MES (w/v) 15-well gel…The gel was then directly scanned at 520 nm for Cy3, 658 nm for Cy5, and 784 nm for Cy7 detection using an Azure Biosystems Sapphire Biomolecular Imager and Azure Spot Analysis Software.

Figure 6. Coagulation factor (APC, thrombin, fXa) labeling in human plasma. (A) Probe concentration optimization assay. Human plasma was
incubated with each fluorescent ABP separately at various probe concentrations ranging from 1 to 20 μM for 60 min at 37 °C. The samples
were then subjected to SDS–PAGE analysis, transferred to a membrane, immunostained with the appropriate antibody, and imaged using an
Azure Biosystems Sapphire Biomolecular Imager as follows: for APC at 658 nm (for Cy5 detection) and 488 nm (for antibody detection), for
thrombin at 784 nm (for Cy7 detection) and 658 nm (for antibody detection), for fXa at 520 nm (for Cy3 detection) and 658 nm (for antibody
detection). The results are representative of at least 3 replicates. (B) Graphical scheme of the methodology used for simultaneous coagulation
factor detection in human plasma. (C) Simultaneous coagulation factor labeling in human plasma. Human plasma was incubated with 5 μM of
each fluorescently labeled ABP and subjected to SDS–PAGE analysis. Direct in-gel analysis was performed with lasers of 520 nm for Cy3, 658
nm for Cy5, and 784 nm for Cy7 using an Azure Biosystems Sapphire Biomolecular Imager. The results are representative of at least 3 replicates.
Figure 6. Coagulation factor (APC, thrombin, fXa) labeling in human plasma. (A) Probe concentration optimization assay. Human plasma was incubated with each fluorescent ABP separately at various probe concentrations ranging from 1 to 20 μM for 60 min at 37 °C. The samples were then subjected to SDS–PAGE analysis, transferred to a membrane, immunostained with the appropriate antibody, and imaged using an Azure Biosystems Sapphire Biomolecular Imager as follows: for APC at 658 nm (for Cy5 detection) and 488 nm (for antibody detection), for thrombin at 784 nm (for Cy7 detection) and 658 nm (for antibody detection), for fXa at 520 nm (for Cy3 detection) and 658 nm (for antibody detection). The results are representative of at least 3 replicates. (B) Graphical scheme of the methodology used for simultaneous coagulation factor detection in human plasma. (C) Simultaneous coagulation factor labeling in human plasma. Human plasma was incubated with 5 μM of each fluorescently labeled ABP and subjected to SDS–PAGE analysis. Direct in-gel analysis was performed with lasers of 520 nm for Cy3, 658 nm for Cy5, and 784 nm for Cy7 using an Azure Biosystems Sapphire Biomolecular Imager. The results are representative of at least 3 replicates.

Method for the elucidation of LAMP products captured on lateral flow strips in a point of care test for HPV 16

Lena Landaverde, Winnie Wong, Gabriela Hernandez, Andy Fan & Catherine Klapperich

…To identify 5′-FAM-specific LAMP products, unstained agarose and acrylamide gels were imaged under 488-nm laser excitation using the Sapphire Biomolecular Imager…

Figure 4. FAM band comparisons of pvuII-digested and undigested lateral flow strip elutions. Three different representations of the LAMP
amplicon products are shown. Gel a and b are LAMP amplicons digested with pvuII (independent of LFS). Gels c, d, f, and g are eluted products
from the LFS itself. Furthermore, gels c and d contain uncleaved LFS-LAMP products (pvuII−), whereas gels f and g are cleaved LFS-LAMP
products (pvuII+). The FAM bands 94–114 (band C) bp and 71–80 bp (b and D) are highlighted as the HPV 16 FAM amplicons. Gel b also shows
the previously sequenced bands of 146 (band A) and 92 (band B). The LFS strips that DNA was eluted from are shown in LFS images e and h
of Fig. 4. The gels in panels a and b are pvuII-digested LAMP products; the lateral flow strips results are equivalent to the LFS shown in panel h.
Figure 4. FAM band comparisons of pvuII-digested and undigested lateral flow strip elutions. Three different representations of the LAMP amplicon products are shown. Gel a and b are LAMP amplicons digested with pvuII (independent of LFS). Gels c, d, f, and g are eluted products from the LFS itself. Furthermore, gels c and d contain uncleaved LFS-LAMP products (pvuII−), whereas gels f and g are cleaved LFS-LAMP products (pvuII+). The FAM bands 94–114 (band C) bp and 71–80 bp (b and D) are highlighted as the HPV 16 FAM amplicons. Gel b also shows the previously sequenced bands of 146 (band A) and 92 (band B). The LFS strips that DNA was eluted from are shown in LFS images e and h of Fig. 4. The gels in panels a and b are pvuII-digested LAMP products; the lateral flow strips results are equivalent to the LFS shown in panel h.

Proteomic Analysis of Endometrial Cancer Tissues from Patients with Type 2 Diabetes Mellitus

Muhammad Mujammami, Mohamed Rafiullah, Assim A. Alfadda, Khalid Akkour, Ibrahim O. Alanazi, Afshan Masood, Mohthash Musambil, Hani Alhalal, Maria Arafah, Anas M. Abdel Rahman and Hicham Benabdelkamel

…The gels were scanned with Sapphire Biomolecular Imager (Azure Biosystems…) and digitalized via the image analysis software Sapphire Capture system (Azure Biosystems…

Figure 1. The representative fluorescent protein of a two-dimensional difference in gel electrophoresis (2D-DIGE) containing tissue samples
from EC Diabetic samples labeled with Cy3 (A), EC Non-Diabetic samples labeled with Cy5 (B), pooled internal control labeled with Cy2 (C),
and merged image (D).
Figure 1. The representative fluorescent protein of a two-dimensional difference in gel electrophoresis (2D-DIGE) containing tissue samples from EC Diabetic samples labeled with Cy3 (A), EC Non-Diabetic samples labeled with Cy5 (B), pooled internal control labeled with Cy2 (C), and merged image (D).
(example of DNA-Protein Binding Assay)

Structure of a RecT/Redβ family recombinase in complex with a duplex intermediate of DNA annealing

Brian J. Caldwell, Andrew S. Norris, Caroline F. Karbowski, Alyssa M. Wiegand, Vicki H. Wysocki & Charles E. Bell

…A gel shift DNA binding assay used two complementary 50-mer oligonucleotides labeled at the 5′-end with either Cy3 or Cy5…For visualization 17.5 μl of each complex was mixed with 7.5 μl Orange G dye…Gels were imaged using a Sapphire Biomolecular Imager (Azure Biosystems) with Sapphire Capture Software (version 1.12.0921.0). Scanning parameters for Fig. 8 were pixel size 100 μm, scan speed high, 2.38mm focus, intensity 2 for Cy5, intensity 4 for Cy3, black lighting 50, white 37186, gamma 1.37. Scanning parameters for Supplementary Fig. 1a, b were intensity 1 for Cy5, intensity 2 for Cy3, black lighting 50, white 15362, gamma 0.88.

Figure 8 (a). Mutational analysis. Each panel shows a gel-shift assay with 3.6 μM of LiRecT mixed with different combinations of Cy3- and Cy5-
labeled 50mer oligonucleotides (25 μM nucleotides). Lanes C3, C5: each oligo without protein. Lanes 3, 5: LiRecT mixed with each individual
oligonucleotide (Cy3-50mer or Cy5-50mer) to form a ssDNA complex. Lanes 35: LiRecT incubated with Cy3- 50mer at 37° for 15 min, followed
by addition of Cy5-50mer and incubation for an additional 15 min to form the duplex intermediate (yellow band).
Figure 8 (a). Mutational analysis. Each panel shows a gel-shift assay with 3.6 μM of LiRecT mixed with different combinations of Cy3- and Cy5- labeled 50mer oligonucleotides (25 μM nucleotides). Lanes C3, C5: each oligo without protein. Lanes 3, 5: LiRecT mixed with each individual oligonucleotide (Cy3-50mer or Cy5-50mer) to form a ssDNA complex. Lanes 35: LiRecT incubated with Cy3- 50mer at 37° for 15 min, followed by addition of Cy5-50mer and incubation for an additional 15 min to form the duplex intermediate (yellow band).
Visible Fluorescence

Neutrophils incite and macrophages avert electrical storm after myocardial infarction

Jana Grune, Andrew J. M. Lewis, Masahiro Yamazoe, Maarten Hulsmans, David Rohde, Ling Xiao, Shuang Zhang, Christiane Ott, David M. Calcagno, Yirong Zhou, Kerstin Timm, Mayooran Shanmuganathan, Fadi E. Pulous, Maximilian J. Schloss, Brody H. Foy, Diane Capen, Claudio Vinegoni, Gregory R. Wojtkiewicz, Yoshiko Iwamoto, Tilman Grune, Dennis Brown, John Higgins, Vanessa M. Ferreira, Neil Herring, Keith M. Channon, Stefan Neubauer, Oxford Acute Myocardial Infarction (OxAMI) Study,* David E. Sosnovik, David J. Milan, Filip K. Swirski, Kevin R. King, Aaron D. Aguirre, Patrick T. Ellinor, and Matthias Nahrendorf

…Oxidative stress was imaged after intravenous injection of CellROX Deep Red Reagent (C10422, Thermo Fisher Scientific, 20 μl diluted in 100 μl of PBS). Hearts were sliced in 1-mm sections for immediate imaging using a Sapphire Biomolecular Imager (Azure Biosystems)…

Figure 3a. Experimental outline. FRI of ROS in hearts 5 hours after MI using the CellROX imaging agent. b, Fluorescence images from cardiac
short axis slices after injection of CellROX. c, Quantification of TBR from FRI. Data are from isotype antibody-injected controls (n = 6 mice) and
neutrophil-depleted mice (n = 7).  Each dot represents a cardiac slice.
Figure 3a. Experimental outline. FRI of ROS in hearts 5 hours after MI using the CellROX imaging agent. b, Fluorescence images from cardiac short axis slices after injection of CellROX. c, Quantification of TBR from FRI. Data are from isotype antibody-injected controls (n = 6 mice) and neutrophil-depleted mice (n = 7). Each dot represents a cardiac slice.
Figure 6m. Representative images of TMRE imaging.
Abbreviations: FRI, fluorescence reflectance imaging; MI, myocardial infarction; ROS, reactive oxygen species; TBR, target-to-background ratio.
Figure 6m. Representative images of TMRE imaging. Abbreviations: FRI, fluorescence reflectance imaging; MI, myocardial infarction; ROS, reactive oxygen species; TBR, target-to-background ratio.
Visible Fluorescence

Eliminating mesothelioma by AAV-vectored, PD1-based vaccination in the tumor microenvironment

Zhiwu Tan, Mei Sum Chiu, Chi Wing Yan, Kwan Man, Zhiwei Chen

…resected REN tumor fluorescence was imaged with a Sapphire Biomolecular Imager (Azure Biosystems) after surgical resection…

Figure 5. Localized injection of rAAV-hsPD1-TWIST1 inhibits human mesothelioma in NSG-huPBL mice. (E) REN tumors in NSG mice (n = 4). 2 x106 REN cells were injected s.c. into NSG mice 3 weeks before i.t. administration of 5 x 1011 g.c. rAAV. Left: tumor growth was measured by bioluminescence imaging. Right: representative tumor fluorescence images at the endpoint. Mock, PBS treatment.
Figure 5. Localized injection of rAAV-hsPD1-TWIST1 inhibits human mesothelioma in NSG-huPBL mice. (E) REN tumors in NSG mice (n = 4). 2 x106 REN cells were injected s.c. into NSG mice 3 weeks before i.t. administration of 5 x 1011 g.c. rAAV. Left: tumor growth was measured by bioluminescence imaging. Right: representative tumor fluorescence images at the endpoint. Mock, PBS treatment.

ZnO Nanowire-Based Early Detection of SARS-CoV-2 Antibody Responses in Asymptomatic Patients with COVID-19

Jung Kim, Sung Kyun Lee, Jong-Hwan Lee, Hye-Yeon Kim, Nam Hoon Kim, Chang Hoon Lee, Chang-Seop Lee, and Hong Gi Kim

…NP antigen in coating buffer A (CB07100, Invitrogen, USA) was prepared at 1, 3, or 5 μg mL−1 and added to the ZnO-NW MP. Next, the anti-NP polyclonal antibody (32, 160, or 800 ng mL−1) in assay buffer (DS98200, Invitrogen, USA) was applied to the plate for 1 h, followed by the Alexa Fluor 488-conjugated secondary antibody (A-11034; 1 mg mL−1) for a further hour. The plate was washed between these steps. The fluorescence signal was measured in a microplate reader and a image taken under a laser scanning imager (Sapphire Biomolecular Imager, Azure biosystems, USA)…

Figure 2 (b). Introduction of SARS-CoV-2 NP antigen onto the surface of the ZnO-NW MP, and detection of anti-SARS-CoV-2 NP IgG polyclonal
antibodies. SARS-CoV-2 NP antigen (1, 3, or 5 μg mL−1) was coated onto the ZnO-NW MP and bare MP. Measurement of the bound rabbit
anti- SARS-CoV-2 NP IgG polyclonal antibody (32, 160, or 800 ng mL−1) using anti-human IgG conjugated to Alexa 488.
Abbreviations: MP, microplate; NP, nucleocapsid; NW, nanowire.
Figure 2 (b). Introduction of SARS-CoV-2 NP antigen onto the surface of the ZnO-NW MP, and detection of anti-SARS-CoV-2 NP IgG polyclonal antibodies. SARS-CoV-2 NP antigen (1, 3, or 5 μg mL−1) was coated onto the ZnO-NW MP and bare MP. Measurement of the bound rabbit anti- SARS-CoV-2 NP IgG polyclonal antibody (32, 160, or 800 ng mL−1) using anti-human IgG conjugated to Alexa 488. Abbreviations: MP, microplate; NP, nucleocapsid; NW, nanowire.

Chemiluminescence Imaging

Shikonin Reduces Growth of Docetaxel-Resistant Prostate Cancer Cells Mainly through Necroptosis

Sascha D. Markowitsch, Kira M. Juetter, Patricia Schupp, Kristine Hauschulte, Olesya Vakhrusheva, Kimberly Sue Slade, Anita Thomas, Igor Tsaur, Jindrich Cinatl, Jr., Martin Michaelis, Thomas Efferth, Axel Haferkamp and Eva Juengel

…To explore the expression and activity of cell cycle and cell death regulating proteins, Western blot analysis was performed…The membranes were incubated with ECL detection reagent (AC2204, Azure Biosystems, Munich, Germany) to visualize proteins with a Sapphire Imager (Azure Biosystems, Munich, Germany)…Cell death regulating proteins were normalized to total protein that was quantified by staining total protein from all membranes with Coomassie brilliant blue and measuring with a Sapphire Imager…
Cancers. 2021;13(4):882
Figure 4. Protein expression profile of cell cycle regulating proteins: Representative Western blot images of cell cycle regulating proteins in
parental (par) and DX-resistant (res) PC3 (left panel) and DU145 (right panel) cells after 48 h exposure to SHI [0.5 µM].
Figure 4. Protein expression profile of cell cycle regulating proteins: Representative Western blot images of cell cycle regulating proteins in parental (par) and DX-resistant (res) PC3 (left panel) and DU145 (right panel) cells after 48 h exposure to SHI [0.5 µM].

NonO Is a Novel Co-factor of PRDM1 and Regulates Inflammatory Response in Monocyte Derived-Dendritic Cells

Kyungwoo Lee, Su Hwa Jang, Hong Tian and Sun Jung Kim
…Proteins bound by antibody were visualized by ECL (Thermo Scientific, #34580 or Advansta, K-12045) and sapphire biomolecular imager (Azure Biosystems)…
Front Immunol. 2020;11:1436
Figure 1(A). Nuclear fraction was immunoprecipitated with anti-PRDM1 antibodies and immunoblotting was performed with anti-PRDM1, Flag-NonO, Flag-TP53BP1, or V5-hnRNPM antibody. A representative image from two independent experiments is shown.
Figure 1(A). Nuclear fraction was immunoprecipitated with anti-PRDM1 antibodies and immunoblotting was performed with anti-PRDM1, Flag-NonO, Flag-TP53BP1, or V5-hnRNPM antibody. A representative image from two independent experiments is shown.

Effects of prolactin on ventricular myocyte shortening and calcium transport in the streptozotocin-induced diabetic rat

Frank C. Howarth, Gunnar Norstedt, Oleksiy I. Boldyriev, Muhammad A. Qureshi, Ozaz Mohamed, Khatija Parekh, Balaji Venkataraman, Sandeep Subramanya, Anatoliy Shmygol, Lina T. Al Kury
…The blots were developed using the Super Signal West Pico Plus chemiluminescent substrate (34577, Thermo Scientific, Rockford, IL, USA). The blot images were acquired using a Sapphire Biomolecular Imager (Azure Biosystems, Dublin, California, USA) using chemiluminescent detection of HRP, coupled with color image acquisition of the protein ladder…
Heliyon. 2020;6(4):e03797
Figure 1. Expression of PRL receptor (PRLR) protein in ventricle tissue from STZ-induced diabetic and control hearts. Typical Western blots
showing expression of PRLR protein in three control (C1 – C3) and 3 diabetic (D1-D3), GAPDH loading control and PRLR protein in tissue from
3 rat mammary glands (MG1-MG3) from 3 female rats, 5 days following delivery.
Figure 1. Expression of PRL receptor (PRLR) protein in ventricle tissue from STZ-induced diabetic and control hearts. Typical Western blots showing expression of PRLR protein in three control (C1 – C3) and 3 diabetic (D1-D3), GAPDH loading control and PRLR protein in tissue from 3 rat mammary glands (MG1-MG3) from 3 female rats, 5 days following delivery.

A dominant negative mitofusin causes mitochondrial perinuclear clusters because of aberrant tethering

Stephanie R Sloat, Suzanne Hoppins
…Membranes were developed in Radiance Plus Chemiluminescent HRP Substrate (Azure Biosystems) for 5 min and imaged on a Sapphire Biomolecular Imager (Azure Biosystems). Band intensities were quantified using AzureSpot analysis software (Azure Biosystems)… Western blot images for shRNA were acquired on Sapphire, and quantification was performed with AzureSpot. Knockdown quantification was normalized using whole-protein stain (Azure Biosystems)
Figure 7. Mfn1S329P is defective for GTP-dependent oligomerization. (A) Representative blue native–PAGE of mitochondria isolated from Flp-In TREx cells expressing Mfn1WT-FLAG or Mfn1S329P-FLAG after incubation with 0.2 μg/ml TET for 4 h. Mitochondria were untreated or incubated in the presence of GMP-PNP (PNP), followed by solubilization and separation by blue native–PAGE and immunoblotted with α-FLAG. Arrow indicates ~200-kD species, closed arrowhead indicates ~320-kD species, and open arrowhead indicates ~450-kD species. Molecular-weight markers are indicated in kD on left. (A, B) Quantification of native mitofusin species indicated in A. Error bars represent mean ± SEM from n = 3 separate experiments.
Figure 7. Mfn1S329P is defective for GTP-dependent oligomerization. (A) Representative blue native–PAGE of mitochondria isolated from Flp-In TREx cells expressing Mfn1WT-FLAG or Mfn1S329P-FLAG after incubation with 0.2 μg/ml TET for 4 h. Mitochondria were untreated or incubated in the presence of GMP-PNP (PNP), followed by solubilization and separation by blue native–PAGE and immunoblotted with α-FLAG. Arrow indicates ~200-kD species, closed arrowhead indicates ~320-kD species, and open arrowhead indicates ~450-kD species. Molecular-weight markers are indicated in kD on left. (A, B) Quantification of native mitofusin species indicated in A. Error bars represent mean ± SEM from n = 3 separate experiments.
Supplemental Figure 6B. Representative
Western blot of α-DHC and α-tubulin in
whole-protein extract from cells treated
with either control shRNA (shLacZ) or
shRNA against DHC (shDHC). Molecularweight markers are indicated in kD on
left. Percent knockdown represents mean
knockdown of DHC n = 3 quantified
by band intensities in Western blot
normalized to whole-protein stain.
Supplemental Figure 6B. Representative Western blot of α-DHC and α-tubulin in whole-protein extract from cells treated with either control shRNA (shLacZ) or shRNA against DHC (shDHC). Molecularweight markers are indicated in kD on left. Percent knockdown represents mean knockdown of DHC n = 3 quantified by band intensities in Western blot normalized to whole-protein stain.

Engineered unnatural ubiquitin for optimal detection of deubiquitinating enzymes

Wioletta Rut, Mikolaj Zmudzinski, Scott J. Snipas, Miklos Bekes, Tony T. Huang and Marcin Drag

…Biotinylated Ub-based probes were detected with a fluorescent streptavidin Alexa Fluor 647 conjugate (1 : 10 000) in TBS-T with 1% BSA, and UCH-L3 was detected with a mouse antihuman monoclonal IgG1 antibody (1 : 1000) and fluorescent goat anti-mouse (1 : 10 000) using an Azure Biosystems Sapphire Biomolecular Imager and Azure Spot Analysis Software.

Chem Sci. 2020;11(23):6058-6069
Figure 5(D). Detection of UCH-L3 in A-431 cell lysates using Ub-based probes (using a streptavidin Alexa Fluor 647 conjugate) and a UCH-L3 antibody.
Figure 5(D). Detection of UCH-L3 in A-431 cell lysates using Ub-based probes (using a streptavidin Alexa Fluor 647 conjugate) and a UCH-L3 antibody.

In vitro Evaluation of ASCs and HUVECs Co-cultures in 3D Biodegradable Hydrogels on Neurite Outgrowth and Vascular Organization

Luís A. Rocha, Eduardo D. Gomes, João L. Afonso, Sara Granja, Fatima Baltazar, Nuno A. Silva, Molly S. Shoichet, Rui A. Sousa, David A. Learmonth and Antonio J. Salgado

…The evaluation of the angiogenic and neurotrophic profile of the previously obtained secretomes was performed using the Human Neuro Discovery Array C1 and Human Angiogenesis Array C1 (RayBiotech, United States)…Finally, the chemiluminescence image of each membrane was obtained using a Sapphire Biomolecular Imager (Azure Biosystems, United States). The intensity of each dot was quantified using the AzureSpot software (Azure Biosystems, United States)…

Figure 5. Analysis of the secretomes of ASCs, HUVECs, and their co-culture after 7 days of culture in GG-GRGDS allowed to understand the
relative expression of a panel of neuroregulatory and angiotrophic molecules. (A) The secretome of ASCs+HUVECs showed an upregulation on
different neurotrophic factors (BDNF, β-NGF, IGF-1, and S-100 B) showing a positive effect of the interaction of both cells on the secretion of
these molecules.
Figure 5. Analysis of the secretomes of ASCs, HUVECs, and their co-culture after 7 days of culture in GG-GRGDS allowed to understand the relative expression of a panel of neuroregulatory and angiotrophic molecules. (A) The secretome of ASCs+HUVECs showed an upregulation on different neurotrophic factors (BDNF, β-NGF, IGF-1, and S-100 B) showing a positive effect of the interaction of both cells on the secretion of these molecules.

A species-specific functional module controls formation of pollen apertures

Byung Ha Lee, Rui Wang, Ingrid M. Moberg, Sarah H. Reeder, Prativa Amom, Michelle H. Tan, Katelyn Amstutz, Pallavi Chandna, Adam Helton, Ekaterina P. Andrianova, Igor B. Zhulin, and Anna A. Dobritsa

Agrobacteria were collected and resuspended in infiltration buffer (10 mM MgCl2, 10 mM MES, 150 μM acetosyringone) at a final concentration of OD600 = 0.8. Pairwise combinations of suspensions were infiltrated into young tobacco leaves, which were then allowed to grow for 3 d in light. A total 12–16 leaves were collected from five to ten plants, the abaxial side of leaves was sprayed with 1 mM luciferin (Biosynth, L-8220) and kept in the dark at 4 °C for 30 min. The bioluminescence images were captured using Azure Sapphire Biomolecule Imager (Azure Biosystems) and converted to heatmaps using the 16-colour look-up table from ImageJ v.1.53a…

Figure 6c. Split-luciferase assay testing the ability of SlINP1 and SlINP2 to interact. Tobacco leaves were divided into sectors co-expressing
indicated proteins containing the N-terminal (NLuc) and C-terminal (CLuc) parts of the firefly luciferase. Panels on the left show the bright-field
images and panels on the right show the corresponding luminescence images. Split-luciferase assay testing the ability of INP1 and INP2 from
Arabidopsis and tomato to interact with a protein from another species. Only the same-species interactions were observed.
Figure 6c. Split-luciferase assay testing the ability of SlINP1 and SlINP2 to interact. Tobacco leaves were divided into sectors co-expressing indicated proteins containing the N-terminal (NLuc) and C-terminal (CLuc) parts of the firefly luciferase. Panels on the left show the bright-field images and panels on the right show the corresponding luminescence images. Split-luciferase assay testing the ability of INP1 and INP2 from Arabidopsis and tomato to interact with a protein from another species. Only the same-species interactions were observed.
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.

NIR Fluorescence Imaging

Accumulation and persistence of ivacaftor in airway epithelia with prolonged treatment

Tara N. Guhr Lee, Deborah M. Cholon, Nancy L. Quinney, Martina Gentzsch, Charles R. Esther Jr

…Blots were probed with mouse monoclonal anti-CFTR antibodies and then with IRDye 680– goat anti-mouse immunoglobulin G (Molecular Probes). Anti-actin (Cell Signaling) was used as a loading control. Protein bands were visualized using a Sapphire Biomolecular Imager (Azure Biosystems)…

Cyst Fibros. 2020;19(5):746-751
Fig. 3 (A). Mature CFTR protein (band C, *) and immature CFTR protein (band B, •) visualized by Western blot analysis of HBE cultures derived from 3 CF (F508del/F508del) patients. Numbers at the top of the lanes represent days since treatment start. Actin is shown as a loading control.
Fig. 3 (A). Mature CFTR protein (band C, *) and immature CFTR protein (band B, •) visualized by Western blot analysis of HBE cultures derived from 3 CF (F508del/F508del) patients. Numbers at the top of the lanes represent days since treatment start. Actin is shown as a loading control.

Differential influence of Streptococcus mitis on host response to metals in reconstructed human skin and oral mucosa

Lin Shang, Dongmei Deng, Sanne Roffel, and Susan Gibbs

tern Blots Fig. 6. Toll-like receptor (TLR) protein expression in reconstructed human skin (RHS) and gingiva (RHG). TLR1 and 4 proteins are shown together with reference tubulin expression. TLR2, 3, 5, and 6 were under the detectable level (data now shown). Data are representative of three independent experiments ee times in PBST d dye-conjugated use (1:7500 for :7500 for TLR1, 2, blots were ecular Imager rnia).…” WESTERN BLOTS Differential influence of Streptococcus …membranes were washed three times in PBST and further incubated with infrared dyeconjugated secondary antibodies against mouse (1:7500 for TLR3, 4, or 5) or against rabbit (1:7500 for TLR1, 2, 6, or tubulin). After washing, the blots were visualized using Sapphire Biomolecular Imager (Azure biosystems, Dublin California)…

Figure 6. Toll-like receptor (TLR) protein expression in reconstructed human skin (RHS) and gingiva (RHG). TLR1 and 4 proteins are shown together with reference tubulin expression. TLR2, 3, 5, and 6 were under the detectable level (data now shown). Data are representative of three independent experiments.
Figure 6. Toll-like receptor (TLR) protein expression in reconstructed human skin (RHS) and gingiva (RHG). TLR1 and 4 proteins are shown together with reference tubulin expression. TLR2, 3, 5, and 6 were under the detectable level (data now shown). Data are representative of three independent experiments.

Alboserpin, the Main Salivary Anticoagulant from the Disease Vector Aedes albopictus, Displays Anti–FXa-PAR Signaling In Vitro and In Vivo

Gaurav Shrivastava, Paola Carolina Valenzuela-Leon, Andrezza Campos Chagas, Olivia Kern, Karina Botello, Yixiang Zhang, Ines Martin-Martin, Markus Berger Oliveira, Lucas Tirloni, and Eric Calvo

…Finally, the plates were scanned at 700 and 800 nm, and the intensity of the labeled proteins was measured using the Azure sapphire biomolecular imager (Azure Biosystems)

Figure 1. Alboserpin inhibits FXa inflammatory effect in vitro. …(E) ERK1/2 protein expression was analyzed using In- Cell Western blots (ICW).
(F) Intensity ratio (p-ERK/ERK).
Figure 1. Alboserpin inhibits FXa inflammatory effect in vitro. …(E) ERK1/2 protein expression was analyzed using In- Cell Western blots (ICW). (F) Intensity ratio (p-ERK/ERK).
Read about On-Cell Western
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.

Phosphor Imaging

High Dose IFN-β Activates GAF to Enhance Expression of ISGF3 Target Genes in MLE12 Epithelial Cells

Kensei Kishimoto, Catera L. Wilder, Justin Buchanan, Minh Nguyen, Chidera Okeke, Alexander Hoffmann and Quen J. Cheng

…Nuclear extracts…were incubated with P32-labaled oligonucleotide probes…The reaction mixtures were run on a 5% acrylamide (30:0.8) gel with 5% glycerol and TGE buffer (24.8 mM Tris, 190 mM glycine, 1 mM EDTA) at 200V for 1 hour and 45 mins. The gels were dried and imaged on a Sapphire Biomolecular imager in phosphor mode (Azure Biosystems, Dublin, CA).

Figure 1. IFN-β Induced STAT1 binds to GAS and
ISRE motifs. (A) EMSA of GAS and ISRE binding
in MLE12 cells treated with IFN-β (100 U/ml). NFY
shown as loading control. Data are representative
of >5 independent experiments.
Figure 1. IFN-β Induced STAT1 binds to GAS and ISRE motifs. (A) EMSA of GAS and ISRE binding in MLE12 cells treated with IFN-β (100 U/ml). NFY shown as loading control. Data are representative of >5 independent experiments.
Figure 5. EMSA of ISRE and GAS binding in response to IFN-β (1 vs.100 U/mll),
IFN-γ (1 vs.100 ng/ml), or a mixture of the low doses. Representative gel from five
replicates.
Figure 5. EMSA of ISRE and GAS binding in response to IFN-β (1 vs.100 U/mll), IFN-γ (1 vs.100 ng/ml), or a mixture of the low doses. Representative gel from five replicates.

Engineered Viral RNA Decay Intermediates to Assess XRN1-mediated decay

Joseph Russo, Cary T. Mundell, Phillida A. Charley, Carol Wilusz, and Jeffrey Wilusz

…The samples were then resolved via denaturing PAGE, dried, exposed to a phosphor screen, and viewed via an Azure Sapphire Biomolecular Imager providing sufficient sensitivity to observe all required bands with one exposure.

Figure 3. In vitro decay assay and xrRNA read-out using different RNA substrates.
Figure 3. In vitro decay assay and xrRNA read-out using different RNA substrates.
Figure 4. Quantification/graphical representation
of gels presented in vitro decay assays and xrRNA
readouts of selected RNA substrates. Using the
representative data obtained in Fig. 3, the relative
levels of the full length input RNA (Input circles) or the
xrRNA reporter decay intermediate (xrRNA squares)
for GAPDH CDS and GAPDH CDS-BNYVV were
quantified by phosphorimaging using pixel counting.
Results are presented as the amount of the indicated
RNA species relative to the band/lane of the gel with
the maximal pixel count for each RNA.
Figure 4. Quantification/graphical representation of gels presented in vitro decay assays and xrRNA readouts of selected RNA substrates. Using the representative data obtained in Fig. 3, the relative levels of the full length input RNA (Input circles) or the xrRNA reporter decay intermediate (xrRNA squares) for GAPDH CDS and GAPDH CDS-BNYVV were quantified by phosphorimaging using pixel counting. Results are presented as the amount of the indicated RNA species relative to the band/lane of the gel with the maximal pixel count for each RNA.

Host cell-dependent late entry step as determinant of hepatitis B virus infection

Xupeng Hong, Yuka Imamura Kawasawa, Stephan Menne, Jianming Hu

…Viral DNAs were resolved on 1.2% agarose gel and detected by 32P-labeled HBV or WHV DNA probes…DNA signals from Southern blot analysis were detected by Sapphire Biomolecular Imager (Azure Biosystems) and quantified using the Image Lab system 6.0.1 (Bio-Rad).

Figure 4. WCH-17 cells were rendered susceptible to HBV infection after huNTCP expression. WCH-17 (A, B) or WC3 (C, D) parental and
huNTCP-expressing cells were plated on regular culture dishes (i.e., with no collagen coating) and infected with ca. 400 genome equivalent
of HBV per cell. Three days post infection, the PF DNA (i.e., Hirt DNA) from mock- or HBV-infected cells was extracted by Hirt extraction and
treated with Exo I/III followed by Southern blot analysis. Hirt DNA from HBV-infected HepG2-huNTCP cells, loaded at 4-fold less than the Hirt
DNA from woodchuck cells, served as the positive control for cccDNA detection.
Figure 4. WCH-17 cells were rendered susceptible to HBV infection after huNTCP expression. WCH-17 (A, B) or WC3 (C, D) parental and huNTCP-expressing cells were plated on regular culture dishes (i.e., with no collagen coating) and infected with ca. 400 genome equivalent of HBV per cell. Three days post infection, the PF DNA (i.e., Hirt DNA) from mock- or HBV-infected cells was extracted by Hirt extraction and treated with Exo I/III followed by Southern blot analysis. Hirt DNA from HBV-infected HepG2-huNTCP cells, loaded at 4-fold less than the Hirt DNA from woodchuck cells, served as the positive control for cccDNA detection.

Preclinical evaluation of [18F]FDG‐PET as a biomarker of lymphoid tissue disease and inflammation in Zika virus infection

Carla Bianca Luena Victorio, Joanne Ong, Jing Yang Tham, Marie Jennifer Reolo, Wisna Novera, Rasha Msallam, Satoru Watanabe, Shirin Kalimuddin, Jenny G. Low, Subhash G. Vasudevan, Ann‐Marie Chacko

…Infected mice were injected i.v. with 10 MBq [18 F]FDG and tissues were harvested following a 60-min tracer uptake. Freshly isolated wholemount lymphoid tissues were immediately exposed to multi-purpose phosphoscreen (BAS-IP MS) for 30 min. Fresh tissues with high tracer uptake were exposed to super-resolution (BAS-IP SR) phosphoscreen for 5 min (GE Healthcare Life Sciences, USA). [ 8 F]FDG standards at 2/3 serial dilution from 600 to 0 kBq were mounted together with mouse tissue for calibration of digital autoradiography (DAR) images. Screens were then scanned using the Sapphire Biomolecular Imager (Azure Biosystems, USA) at 100-μm resolution…

Figure 3. Ex vivo assessments of tissue [18F]FDG uptake in acute ZIKV and DENV disease. a, b Representative wholemount tissue ex vivo digital
autora-diography (DAR) images of a lymphoid tissues, testes, and liver and b brain, heart, and digestive tract from pre-infection (n = 5), late
ZIKV (n = 6), and late DENV (n = 8) mice.
Abbreviations: DENV, dengue virus; ZIKV, Zika virus
Figure 3. Ex vivo assessments of tissue [18F]FDG uptake in acute ZIKV and DENV disease. a, b Representative wholemount tissue ex vivo digital autora-diography (DAR) images of a lymphoid tissues, testes, and liver and b brain, heart, and digestive tract from pre-infection (n = 5), late ZIKV (n = 6), and late DENV (n = 8) mice. Abbreviations: DENV, dengue virus; ZIKV, Zika virus

Identification and functional analysis of a galactosyltransferase capable of cholesterol glycolipid formation in the Lyme disease spirochete Borrelia burgdorferi

Petronella R. Hove, Forgivemore Magunda, Maria Angela de Mello Marques, M. Nurul Islam, Marisa R. Harton, Mary Jackson, John T. Belisle

…The lipid extracts were resolved by thin-layer chromatography (TLC) using silica gel G60 TLC plates and chloroform/methanol (90:10 v/v) as the mobile phase. Reference non-radioactive standards were resolved concurrently and developed separately from the blot containing radioactive lipids with phosphomolybdic acid stain. Following this, blot with reference standard was matched with TLC blot to mark resolution of lipids. After development radiolabeled lipids were imaged using Azure Sapphire Biomolecular Imager (Azure Biosystems Inc, Dublin, CA)…

Figure 5. TLC of lipids from cell free assays for the
enzymatic incorporation of [26- 14C] cholesterol into
cholesteryl-β-D-galacto-pyranoside (CGal). A. The
formation of cholesteryl-β-D-galactopyranoside (CGal)
(arrow) in whole cell lysates of Bb and recombinant E.
coli expressing WT bb0572 (lane 1 and 2). As expected,
CGal was not formed by the whole cell lysate of E. coli
expressing mutant bb0572 ΔCFF/I/DGD (lane 3). Lane 4
is the empty expression vector control, and lane 6 and 7
are from boiled whole cell lysates of Bb and recombinant
E. coli expressing WT bb0572, respectively. Note lane 5
was not used. B. Substrate controls: Bb and E. coli WT
bb0572 whole cell lysates with UDP-Gal (lane 1 and 2);
GDP-Man (lanes 3 and 4), or cholesterol oxidase treated
[26- 14C] cholesterol with UDP-Gal (lane 5 and 6) as
substrate. CHO, cholesterol; ACGal, cholesteryl 6-O-acylβ-D-galactopyranoside and MGal, monogalactosyl
diacylglycerol. Ec WT (Escherichia coli wild type), Ec MT
(E.coli mutant—bb0572 ΔCFF/DGD/I) and Ec VC- E.coli
vector control).
Figure 5. TLC of lipids from cell free assays for the enzymatic incorporation of [26- 14C] cholesterol into cholesteryl-β-D-galacto-pyranoside (CGal). A. The formation of cholesteryl-β-D-galactopyranoside (CGal) (arrow) in whole cell lysates of Bb and recombinant E. coli expressing WT bb0572 (lane 1 and 2). As expected, CGal was not formed by the whole cell lysate of E. coli expressing mutant bb0572 ΔCFF/I/DGD (lane 3). Lane 4 is the empty expression vector control, and lane 6 and 7 are from boiled whole cell lysates of Bb and recombinant E. coli expressing WT bb0572, respectively. Note lane 5 was not used. B. Substrate controls: Bb and E. coli WT bb0572 whole cell lysates with UDP-Gal (lane 1 and 2); GDP-Man (lanes 3 and 4), or cholesterol oxidase treated [26- 14C] cholesterol with UDP-Gal (lane 5 and 6) as substrate. CHO, cholesterol; ACGal, cholesteryl 6-O-acylβ-D-galactopyranoside and MGal, monogalactosyl diacylglycerol. Ec WT (Escherichia coli wild type), Ec MT (E.coli mutant—bb0572 ΔCFF/DGD/I) and Ec VC- E.coli vector control).
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.

White Light Imaging

A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)

Jong-Hwan Lee, Minsuk Choi, Yujin Jung, Sung Kyun Lee, Chang-Seop Lee, Jung Kim, Jongwoo Kim, Nam Hoon Kim, Bum-Tae Kim, Hong Gi Kim

…the intensity of the test and control lines were converted to peak histograms using a Sapphire Biomolecular Imager.

Figure 4. Identification of the sandwich pair for detection of
SARS-CoV-2 spike antigen. a) Schematic diagram of LFIA
using ACE2 as the capture probe and sandwich analysis
results obtained from paired antibodies (CR3022, F26G19,
and S1mAb). SARS-CoV-2 S1 antigen (50 ng) was used as a
positive control, and buffer containing no S1 antigen was used
as a negative control. After 20 min, the strips were captured
by a smartphone, and their peak intensities were analyzed. b)
Schematic diagram of LFIA, using antibodies as the capture
probe, and their sandwich analysis results. c) Peak intensities
of capture probe (PC)–detection probe (PD) pairs. A total of 12
pairs of positive controls (50 ng S1 antigen) were tested, and
their intensities were analyzed. Peak intensity was calculated
by subtracting the background intensity of the strip from the
average intensities of the dots.
Figure 4. Identification of the sandwich pair for detection of SARS-CoV-2 spike antigen. a) Schematic diagram of LFIA using ACE2 as the capture probe and sandwich analysis results obtained from paired antibodies (CR3022, F26G19, and S1mAb). SARS-CoV-2 S1 antigen (50 ng) was used as a positive control, and buffer containing no S1 antigen was used as a negative control. After 20 min, the strips were captured by a smartphone, and their peak intensities were analyzed. b) Schematic diagram of LFIA, using antibodies as the capture probe, and their sandwich analysis results. c) Peak intensities of capture probe (PC)–detection probe (PD) pairs. A total of 12 pairs of positive controls (50 ng S1 antigen) were tested, and their intensities were analyzed. Peak intensity was calculated by subtracting the background intensity of the strip from the average intensities of the dots.
Figure 5a. Sensitivity and specificity of the ACE2-based LFA.
a) Results of ACE2-based LFA for the detection sensitivity of
SARS-CoV-2 S1 antigen. Serially diluted antigen concentrates
(concentration range: 500 ng/mL to 5 ng/mL) were tested
by ACE2-based LFA. After 20 min, the LFA strips were
photographed with a smartphone. Moreover, the intensity of
the test and control lines was converted to a peak histogram
by an image analyzer.
Figure 5a. Sensitivity and specificity of the ACE2-based LFA. a) Results of ACE2-based LFA for the detection sensitivity of SARS-CoV-2 S1 antigen. Serially diluted antigen concentrates (concentration range: 500 ng/mL to 5 ng/mL) were tested by ACE2-based LFA. After 20 min, the LFA strips were photographed with a smartphone. Moreover, the intensity of the test and control lines was converted to a peak histogram by an image analyzer.
Figure 6b. Laboratory confirmation of ACE2-based LFIA
using clinical samples. b) Results of ACE2-based LFA for the
detection sensitivity of cultured SARS-CoV-2. Serially diluted
virus concentrates (concentration range: 1.07 × 108 copies/
mL to 5.35 × 106 copies/mL) were tested. After 20 min, the
LFIA strip was taken with a smartphone and scanned with an
image analyzer. The line intensities of the test and control lines
were converted to peak histograms. Also, the intensity of the
test lines was measured by a portable line analyzer (IL: line
intensity of test line). Furthermore, human coronavirus (OC43)
was tested as a negative control.
Figure 6b. Laboratory confirmation of ACE2-based LFIA using clinical samples. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 108 copies/ mL to 5.35 × 106 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (IL: line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control.

A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)

Jong-Hwan Lee, Minsuk Choi, Yujin Jung, Sung Kyun Lee, Chang-Seop Lee, Jung Kim, Jongwoo Kim, Nam Hoon Kim, Bum-Tae Kim, Hong Gi Kim

…The clonogenic recovery potential gives insight into the capability of the cells to form a new tumor (metastasis). Therefore, 500 cells/well were seeded on a 6-well-plate and treated for 10 days with ART. Untreated cells served as controls. RCC cells were subsequently fixed with 85% MeOH/15% AcOH and stained with Coomassie (0.5 g Coomassie Blue G250 (SigmaAldrich, Darmstadt, Germany), 75 mL AcOH, 200 mL MeOH, 725 mL distilled water). Amount and size of cell clone colonies were measured with a biomolecular imager (Sapphire, Azure Biosystems, Biozym, Hess. Oldendorf, Germany)…

Figure 3. Clonogenic growth of RCC cells: Clonogenic growth of parental and resistant Caki-1 (a), 786-O (b), KTCTL-26 (c), and A 498 (d)
cells treated with ART (10–50 μM) for 10 days. Untreated cells served as controls (set to 100%). Error bars indicate standard deviation (SD).
Significant difference to untreated control: ** p ≤ 0.01, *** p ≤ 0.001. n = 5.
Figure 3. Clonogenic growth of RCC cells: Clonogenic growth of parental and resistant Caki-1 (a), 786-O (b), KTCTL-26 (c), and A 498 (d) cells treated with ART (10–50 μM) for 10 days. Untreated cells served as controls (set to 100%). Error bars indicate standard deviation (SD). Significant difference to untreated control: ** p ≤ 0.01, *** p ≤ 0.001. n = 5.
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.
Lorem ipsum dolor sit amet, consectetur adipisicing elit. Optio, neque qui velit. Magni dolorum quidem ipsam eligendi, totam, facilis laudantium cum accusamus ullam voluptatibus commodi numquam, error, est. Ea, consequatur.

Applications

Recently Published

Azure Imagers have been published in over 2,000 publications worldwide. Our high-resolution CCD cameras are the reason behind reproducible data and publication-ready images.

Impressed with the Sapphire FL and want to quickly share relevant publications with a colleague? Download a free booklet with publications featuring the SFL here.

Specifications

ParameterAzure Sapphire FL
Detection modesFluorescence, phosphor imaging, densitometry, and chemiluminescence*
DetectorsConfocal scanning with lasers and PMT or APD detectors
Pixel size5, 10, 20, 25, 50, 75, 100, 150, 200, 500, 1000 μm
Supported wavelength range375 – 900 nm
Excitation range375 – 850 nm
Emission range380 – 900 nm
Maximum field of view (FOV):25 cm x 25 cm
Z-plane focal range7 mm; minimum delta 0.01mm, from -1 mm below the glass to 6 mm above the glass
Standard optical modulesSolid-state laser diodes emitting at 375, 488, 532, 638, 658, 685, and 784 nm. Phosphor Standard Optical Module available.
Custom Optical ModulesCustom lasers and filters are available upon request to provide exceptional research flexibility.
Wide dynamic range24-bit
Clearance from platen to lid4 cm
Animal imagingYes, with 5 built-in anesthesia ports compatible with standard small animal anesthesia tubing
Unit dimensions (W x H x D)​63.0 cm x 39.9 cm x 59.3 cm / 24.8” x 15.7” x 23.3"
Weight​43.5 kg / 95.9 lbs
Power requirements100 – 240 VAC ± 10%, 50/60 Hz
Connection to computerFast LAN (Ethernet) cable
CertificationsCE, cTUVus, CB scheme

*Chemiluminescence module sold separately

Resources

Document TypeDescription
VideoHow the Sapphire FL Supports In Vitro to In Vivo ResearchWATCH
BrochureAzure Sapphire FL Biomolecular Imager BrochureDOWNLOAD
BookletSapphire FL Biomolecular Imager Applications and PublicationsDOWNLOAD
User ManualSapphire FL User ManualDOWNLOAD
App NoteFluorescent IHC Imaging of Tumor Tissue Arrays on the Sapphire FLDOWNLOAD
App NoteIn vivo imaging and monitoring of RFP+ tumors in mice using the Sapphire FLDOWNLOAD
App NoteMulticolor Fluorescent Imaging of Cultured Cells on the Sapphire FL: Live/Dead (AO/PI) AssaysDOWNLOAD
App NoteSapphire FL’s Laser and Filter Flexibility Allows Imaging of Dyes with Large Stokes ShiftsDOWNLOAD
App NoteUsing Total Cell Staining and Advanced Imaging to Unlock the Full Power of In-Cell WesternsDOWNLOAD
App NoteImaging YFP-expressing cells with the Sapphire FLDOWNLOAD
App NoteMultiplex Fluorescent Immunohistochemistry SlidesDOWNLOAD
App NoteBiFC Assay for Determination of in-vitro Protein-Ligand Interaction using the SapphireDOWNLOAD
App NotePhosphor Imaging with the Sapphire Biomolecular ImagerDOWNLOAD
App NoteDetecting Proteins In-Situ with In-Cell Western BlottingDOWNLOAD
App NoteIncreasing Assay Efficiency with Four Color DetectionDOWNLOAD
FlyerHow In-Cell Westerns Can Advance Your ResearchDOWNLOAD

Shop Sapphire FL Accessories

Sapphire FL Pricing

See what the hype is about. Let's chat.

What Scientists Say about the Sapphire FL

Our lab recently acquired a Sapphire FL Biomolecular Imager. This imager is a game-changer, and the analysis software is fantastic. Going from regularly using an imager built in the 90’s to a unit this modern is a substantial leap forward, but this overdue upgrade in machinery is not the sole reason for our excitement. Its sensitivity, resolution, and user-friendly UI speak volumes for the quality of the product and has made imaging an even more enjoyable part of our experiments. My colleagues and I are looking forward to using it for years to come… more

Ryan Steere
Ryan Steere
Graduate Student, Ohio University
Shopping cart0
There are no products in the cart!
Continue shopping