A low-cost method for visible fluorescence imaging
| Autor: | Marc L. Pusey, Crissy L. Tarver |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Fluorescence-lifetime imaging microscopy protein crystallization Biophysics Analytical chemistry Color Lactoglobulins Biochemistry Fluorescence Research Communications law.invention Crystal 03 medical and health sciences symbols.namesake Structural Biology law Stokes shift Microscopy Concanavalin A Genetics Crystallization Fluorescent Dyes biology Rhodamines Chemistry Proteins Equipment Design Condensed Matter Physics Molecular Imaging 3. Good health 030104 developmental biology Costs and Cost Analysis symbols biology.protein Smartphone trace fluorescent labeling low-cost imaging Protein crystallization |
| Zdroj: | Acta Crystallographica. Section F, Structural Biology Communications |
| ISSN: | 2053-230X |
| DOI: | 10.1107/s2053230x17015941 |
| Popis: | A very low cost method for carrying out imaging of trace fluorescently labeled protein crystallization plates is described. A wide variety of crystallization solutions are screened to establish conditions that promote the growth of a diffraction-quality crystal. Screening these conditions requires the assessment of many crystallization plates for the presence of crystals. Automated systems for screening and imaging are very expensive. A simple approach to imaging trace fluorescently labeled protein crystals in crystallization plates has been devised, and can be implemented at a cost as low as $50. The proteins β-lactoglobulin B, trypsin and purified concanavalin A (ConA) were trace fluorescently labeled using three different fluorescent probes: Cascade Yellow (CY), Carboxyrhodamine 6G (CR) and Pacific Blue (PB). A crystallization screening plate was set up using β-lactoglobulin B labeled with CR, trypsin labeled with CY, ConA labeled with each probe, and a mixture consisting of 50% PB-labeled ConA and 50% CR-labeled ConA. The wells of these plates were imaged using a commercially available macro-imaging lens attachment for smart devices that have a camera. Several types of macro lens attachments were tested with smartphones and tablets. Images with the highest quality were obtained with an iPhone 6S and an AUKEY Ora 10× macro lens. Depending upon the fluorescent probe employed and its Stokes shift, a light-emitting diode or a laser diode was used for excitation. An emission filter was used for the imaging of protein crystals labeled with CR and crystals with two-color fluorescence. This approach can also be used with microscopy systems commonly used to observe crystallization plates. |
| Databáze: | OpenAIRE |
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