Creating Biomedical Technologies to Improve Health

LABS AT NIBIB

High throughput single molecule imaging microscope

The high throughput single molecule microscope was purpose designed to image large numbers of cell samples in a multi-well format. This microscope can routinely image up to 8 coverslips and 10,000+ cells per acquisition. An included analysis pipeline automatically segments images to identify nuclei, cell bodies, and puncta (RNAs, receptors, etc.), and generates statistics on these puncta (number/cell, co-expression, co-localization, etc.). This microscope is currently located in the Larson Lab and will move to AIM once the facility is operational.

Donating lab: Daniel Larson, NCI

Spatial resolution: 240nm in X, Y. 930nm in Z.

Temporal resolution: 200Hz framerate.

Excitation wavelengths: Seven Lumencor (LED) excitation bands - violet, blue, cyan, teal, green, yellow and red.

Detection channels: One sCMOS camera. Multiple emission bands are acquired sequentially. Currently available emission bands include DAPI, GFP, Cy3, Cy5, or similar fluorophores. Other emission bands can be easily accommodated.   

Maximum field of view (1 z-stack): 330 microns x 330 microns

Objectives: 40X 1.4NA Oil (Zeiss)

Sample specifications: Samples are imaged through #1.5 (0.17mm thickness) glass coverslips in a standard inverted microscope geometry. The microscope can be programmed to automatically acquire images from multiple coverslips or multi-well plates. Field of view is approximately 330 microns. This is a widefield, epifluorescence imaging set-up and as such has no background rejection, making it ideally suited for thin samples (<20 microns) such as cultured cells.