Using Auto Contrast in Volocity

I’ve had a lot of questions recently concerning the “auto-contrast” feature of Volocity Acquisition. So I thought I would put down some information here to refer people to in the future. Essentially the auto-contrast allows you to image very faint signals and still see them with relatively short exposure times. This can be useful in Read more about Using Auto Contrast in Volocity[…]

OMX Sample Preparation

To achieve best results using structured illumination to provide 2-fold improvement in resolution requires that samples be prepared carefully. Read the document below which provides some guidelines for preparing your samples to image in the University of Alberta’s OMX. Also see www.omx.med.ualberta.ca for more details. OMX Sample Preparation Guidelines

STORM Immunofluorescence Protocol

Solutions Fixative: 3% paraformaldehyde + 0.1% glutaraldehyde in PBS) Fixative Quench: 0.1% Sodium borohydride (NaBH4) in PBS Blocking Buffer (BB): 3% BSA, 0.2% Triton X-100 in PBS Washing Buffer (WB): 0.2% BSA, 0.1% Triton X-100 in PBS Imaging buffers: 10 % Glucose solution in PBS Oxygen scavenger system: Final: 0.5 mg/ml glucose oxidase (Sigma G2133). Read more about STORM Immunofluorescence Protocol[…]

Sample Preparation for STED Microscopy

Please download the following pdf file and read it before attempting any immunofluorescence protocols to be used on the STED microscope. Leica will be shipping some of the specially required fluorochromes conjugated to ant-mouse and anti-rabbit antibodies, so if you have a requirement for an antibody against an unusual primary species please let me know Read more about Sample Preparation for STED Microscopy[…]

Count Nuclei (or other spots) in an image

Counting Nuclei (or other features) in an image is a common analysis problem. A very common biological sample for microscopy is DAPI stained DNA in cell nuclei. The staining delineates the nuclei pretty well, because in a metaphase cell there is DNA all over the nucleus. however, the staining is not homogenous, as there are Read more about Count Nuclei (or other spots) in an image[…]

Preparing a MATERIALS & METHODS section

(This is based in large part on a page from the Nikon Imaging Centre at Harvard Medical School, https://nic.med.harvard.edu/equipment/materialsandmethods)   Here’s what to include in a MATERIALS AND METHODS section for the microscopy section. For a reader to assess whether you have performed experiments correctly, there are a few parameters that you need to include in Read more about Preparing a MATERIALS & METHODS section[…]