One of the most common problems with see with researchers is that they struggle with optimizing antibody labeling for their specific application. Typically, what we see most commonly is that researchers are unable to achieve uniform penetration of their antibody across the depth of a tissues. Below we provide steps for how best to optimize antibody labeling to save both time and month. However, there are are a few key considerations that should be evaluated before proceeding with antibody labeling.
- Minimize the thickness of the tissue that you are trying to label. A 2 mm thick tissue will take 3-4 times longer to label than a 1 mm thick tissue and might not be able to be imaged with your microscope system.
- Use secondary antibodies to enhance your signal. This will increase the processing time but will greatly improve your results.
- Start small and work your way up! This is the most important consideration in adopting tissue clearing as some antibodies will not penetrate tissues well at all. Before you spend weeks of time and thousands of dollars in reagents, start small with thin tissues and work your way up as you optimize.
With the Visikol HISTO tissue clearing approach you are now able to rapidly and easily image tissues in 3D without damaging the underlying cellular morphology. This means that you can characterize all of the cells within your 3D cell culture models instead of just the periphery.
The Visikol HISTO approach has been designed to pair with immunolabeling or fluorescent protein and is reversible so that 3D histology can be followed up with traditional 2D histology. The Visikol HISTO-M approach is designed specifically for use with 3D cell cultures (e.g. organoids, microtissues, spheroids) and high-throughput imaging as it is compatible with well plates and liquid handling robots.
Visikol HISTO-M has a refractive index of 1.48 and approx. 0.1 mL is required per well in a 96 well plate.
Refractive Index: 1.48
Composition: Organic solvent based
Visikol HISTO-M can be stored indefinitely at room temperature away from direct sunlight.