Widefield microscopy generally refers to the use of a microscope not equipped with a pinhole (see Confocal Microscopy) in the detection path. These microscopes can be used for capturing both transmitted light images (phase contrast, DIC, etc.) as well as fluorescent images. In this instrument a light source (mercury bulb, LED, etc.) is used to evenly illuminate your field of view all at once and excite fluorescent probes with which the imaged sample is labeled. The emitted fluorescence is captured using a camera. Widefield microscopy is an excellent choice for relatively “thin” samples (tissue culture, cryosections, etc.) where the samples are less than 10 microns.
Modern widefield microscopes support 4 and more color channels and are well-suited for multicolor imaging. They are capable of live-cell imaging but for particularly light-sensitive applications, other techniques should be considered.
Because this instrument does not have any way to eliminate out of focus light, samples thicker than 10 microns may have dramatically reduced image quality. When working with thicker samples, Confocal Microscopy may be a better choice.
Availability at Yale
Yale University has a large number of widefield microscopes. Facilities that provide access to widefield microscopes include:
- CCMI – confocal microscopy, 2-photon microscopy, STED super-resolution microscopy
Please contact us if you think your research in hematology might benefit from using these kind of imaging techniques. We are happy to meet with you, discuss your particular application with you and explore available options. We also offer limited financial support that will allow researchers at all career stages to try out new imaging techniques without any costs.