Azure Biosystems
Counting bacterial colonies is a great means of identifying the general bacteria content of a sample. Growing bacteria cultures on a gelled-growth medium, like an agar plate or petri dish, permits researchers identify colonial characteristics. Bacteria or other microorganism colonies can be observed by scientists through various staining and imaging methods. The process of imaging bacterial colonies is made simple by using gel imagers, such as the Azure Imaging Systems.
Capturing fluorophores that fluoresce in the visible spectrum is useful many applications. Here are some examples of when capturing fluorophores might be useful:
Crystal violet is used to measure cell viability of adherent cells. Using crystal violet is an easy and efficient method for the quantification of cell density in cell culture plates.
During the crystal violet assay, dead cells are washed away and the remaining attached live cells are visualized with the crystal violet dye. This assay method works by the utilization of protein-DNA binding that results in the staining of the nucleus dark blue and the cytoplasm light blue. Crystal violet absorbs at 595 nm.
The crystal violet assay process occurs quickly, providing fast results and reliable readings. With this assay, it is possible to determine the number of cells in a culture, as well as their concentration, characteristics, and behavior. It is a highly recommended technique for researchers and scientists who depend on the analysis of cell culture density in their work.
By using crystal violet assay, researchers can save time and resources, while also obtaining highly accurate data that can further their understanding of various biological processes and phenomena.
GFP stands for green fluorescent protein. GFP is a protein produced by the bioluminescent jellyfish Aequorea victoria (also known as “crystal jelly”). As its name suggests, GFP emits bioluminescence in the green zone of the visible spectrum. In modern day, GFP is cloned and commonly used in molecular biology as fluorescent markers of gene expression. It emits bright green light (lambda max = 510 nm) when excited with ultraviolet (UV). GFP is expressed in many species, including yeasts, bacterium, fish, fungi, and plants. A dish of GFP-expressing marijuana is shown in Figure 1.
An advantage of GFP expression is that it is highly sensitive and thus can be used to visualize primary cellular functions, such as signal transduction, protein translation, and DNA replication.
Imaging bacterial colonies can be done using laser scanners, like the new Azure Sapphire FL Biomolecular Imager, or Western blot imaging systems, like the Azure Imaging Systems. The Sapphire FL enables imaging and quantitation of several multi-well plates at a time with a 25x25cm scanning area, ensuring the absorbance of each well easily measured. Azure Imagers can also be used to image more than just gels and membranes.
Because of the depth of field, the Azure Imagers can capture images of bacterial or yeast colonies on plates. Every Azure Imager can capture white light images, but only the Azure 400 and 600 Systems can image fluorophores that fluoresce in the visible spectrum. These two imagers allow you to capture of microorganisms expressing fluorophores, such as GFP (Figure 1), mCherry (Figure 2), or both (Figure 3).
Other sources of fluorescence on a flat surface can be captured by imagers, including bacterial colonies on a plate. Fluorescent proteins, like GFP variants, fall in the visible range. Digital imagers, like the Azure 400, can be used for more than just gels and membranes. The Azure 600 is a great for image bacterial colonies on plates because it offers two-channel NIR fluorescent detection.
There are multiple analysis software options available on the market to count bacterial colonies. AzureSpot Pro is an in-depth analysis software that includes a Colony Counter module, making the process of counting of spots or features fast and easy. Download a free trial of AzureSpot Pro by clicking here.
Analysis software for the analysis of plates, gels, cell-based assays, and more
All-around application ace: NIR, RGB, chemi, blue light, white light, and UV
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Western blotting is a widely used analytical technique that can identify one or more specific proteins in a complex mixture of proteins. It is a
We’ve all been there… Non-specific bands are a perennially frustrating problem in Western blotting. Here are some of the different reasons you might be getting
Chemiluminescent detection (Figure 1) depends on an enzymatic reaction so timing and the amount of both enzyme and substrate used have important effects on data quality.
