Mitochondria imaged with the ReScan Confocal (RCM) microscope




Microscope setup

Nikon Ti2 with Okolabs incubator and PFS (Perfect Focus System)

CFI Plan Fluor 40x oil 1.3 objective

ReScan Confocal add-on (RCM) from

Omicron 561nm 20mW laser

Hamamatsu Orca Flash 4.0 V3 camera 


Acquisition details

Framerate: 1 frame per 10 seconds

Duration: 61h06m

2048*2044 pixels

Laserpower: 1 microwatt at sample plane

Pixel dwell time: about 1 microsecond


Sample details

Cell type: HO1N1 cells 

Staining: Bacmam 2.0 Mitochrondria RFP (Invitrogen™ C10601)

Glass bottom 8-well IBIDI chamber 

Prolong Live 1:100 (LIFE TECHNOLOGIES P36975)


Cost of microscope 

ReScan Confocal add-on: <40k EUR

Cost of label: 10 EUR



As the movie clearly illustrates there is no sign of phototoxicity. Mitochondria are very mobile during the entire timelapse. Also mitochondrial fusion and fission can be observed. The captured cells undergo mitosis at the start of the movie, indicating they tolerate the imaging very well. 

The observed decay in fluorescence intensity can be attributed to the fact that the labeling method is based on the Bacmam expression system. With each cell division the number of viral particles is approximately halved, therefore lowering expression.

This proves the strong point of the ReScan Confocal (RCM) add-on: sensitivity. Because of its unique camera based detection very low excitation energies can be used and still high signal to noise is preserved.



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