26/05/2013 1 Microscopia in fluorescenza BUSB http://www.zeiss.de/C12567BE0045ACF1/Contents-Frame/10BBB27317CDCE32C125752000466B8F Fluorocromi comuni Fluorochromes are stains, somewhat similar to the better- known tissue stains, which attach themselves to visible or sub-visible organic matter. These fluorochromes, capable of absorbing and then re-radiating light, are often highly specific in their attachment targeting and have significant yield in absorption- emission ratios. Absorbance ln (I o /I) = σnd (Beer –Lambert law) I o = light intensity entering cuvette I=light intensity leaving cuvette σ : absorption cross section n molecules d = cross section (cm) or ln (I o /I) = α C d (Beer –Lambert law) a=absorption coefficient C = concentration • Converting to decimal logs and standardizing quantities we get Log (I 0 /I) = εcd = A Now e is the decadic molar extinction coefficient A = absorbance or optical density (OD) a dimensionless quantity d n molecules σ – absorption cross section
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Microscopia in fluorescenza - unipv · (campo chiaro) Light Sources ... (microscopio confocale) Microscopio in fluorescenza ... Percorso ottico dell’eccitazione e
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Fluorochromes are stains, somewhat similar to the better-known tissue stains, which attach themselves to visible or sub-visible organic matter. These fluorochromes, capable of absorbing and then re-radiating light, are often highly specific in their attachment targeting and have significant yield in absorption-emission ratios.
Immunofluorescence analysis of mesenchymal stem cells using Zymed Ms anti-Stro-1 (Cat. No. 39-8401) and Gt anti-Mouse-Alexa Fluor 488 (Cat. No. A21042) (green). Tubulin is stained with phalloidin-Alexa 594 (red) and nuclei are stained with DAPI (blue). Sample is mounted in ProLong Gold antifade reagent.
Fluorocromizzazione del DNA con Ioduro di Propidio
Fluorescence Microscope withColor Video (CCD) 35 mm Camera
Cameras and emission filters
Color CCD camera does not need optical filters to collect all wavelengths but if you want to collect each emission wavelength optimally, you need a monochrome camera with separate emission filters shown on the right (camera is not in position in this photo).