MacBiophotonics

 

 

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Microscopy - Ratio Imaging

  • Calcium imaging (fura-red, fura2, pericams)

  • Redox (roGFP)

  • pH imaging (SNARF, SNAFL, BCECF)

  • FRET-based biosensors (CFP-YFP)

  • Confocal or widefield

System details

Microscope: Zeiss Axiovert 100.

Excitation: Sutter DG4 fast (1.2 ms between positions) excitation switcher.

CARV2 spinning disc confocal scanhead. Optional confocal or widefield imaging.

Emission: Ludl MAC5000 filter wheel

Objectives: 10×; 40× 1.3NA FLUOR oil ; 100× 1.4NA oil plan apo

Camera: Cascade 2 1024B EMCCD .

Acquisition Software: IPLab

Ratio-imaging timecourse of HeLa cells loaded with Fura red and treated with histamine to evoke cytoplasmic calcium oscillations.

Filter sets 

RFP: Semrock TRITC-A-000. 543/22; 562 dichroic; 593/40.

GFP: Semrock 3035B. 472/30; 495 dichroic; 520/35.

DAPI: Semrock 5060B. 377/50; 409 dichroic; 447/60.

 

Fura2: Chroma D340xv2; D380xv2; 505DCXRU; HQ535/50m.

Ratiometric-Pericam: Chroma D414/30x; HQ485/15x; 505DCXRU; HQ535/50m.

Fura Red: emission 650/40nm

SNAFL: dual excitation set.

CFP-YFP dual set.

 

Separate excitation filters for DG4.

DAPI: Semrock BrightLine 377/50.

CFP: Semrock BrightLine 427/10.

GFP: Semrock BrightLine 482/35.

YFP: Semrock BrightLine 504/12.

 

 

 

 

 
 

Updated 08/17/2009

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