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Versatile, do-it-yourself, low-cost spinning disk confocal microscope

Aaron R. Halpern, Min Yen Lee, Marco D. Howard, Marcus A. Woodworth, Philip R. Nicovich, Joshua C. Vaughan

2022Biomedical Optics Express32 citationsDOIOpen Access PDF

Abstract

Confocal microscopy is an invaluable tool for 3D imaging of biological specimens, however, accessibility is often limited to core facilities due to the high cost of the hardware. We describe an inexpensive do-it-yourself (DIY) spinning disk confocal microscope (SDCM) module based on a commercially fabricated chromium photomask that can be added on to a laser-illuminated epifluorescence microscope. The SDCM achieves strong performance across a wide wavelength range (∼400-800 nm) as demonstrated through a series of biological imaging applications that include conventional microscopy (immunofluorescence, small-molecule stains, and fluorescence in situ hybridization) and super-resolution microscopy (single-molecule localization microscopy and expansion microscopy). This low-cost and simple DIY SDCM is well-documented and should help increase accessibility to confocal microscopy for researchers.

Topics & Concepts

ConfocalMicroscopeConfocal microscopyMicroscopyMaterials scienceFluorescence microscopeOpticsLight sheet fluorescence microscopyBiological imagingPhotomaskOptical microscopeScanning confocal electron microscopySuper-resolution microscopySpinningFluorescence-lifetime imaging microscopyBright-field microscopyOptical sectioningBiological specimen4Pi microscopeNanotechnologyFluorescenceImage processingPoint spread functionAdvanced Fluorescence Microscopy TechniquesAdvanced optical system designRandom lasers and scattering media
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