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Klinik für Innere Medizin IHämatologie, Onkologie und Stammzelltransplantation

Microscopy

Olympus Scan^R

Thanks to the generous support of the Medical Faculty, the Olympus scanR system has just undergone a major update/upgrade, which also includes the new AI analysis module from Olympus. More details will be posted soon.

Lighthouse has an Olympus Scan^R High Content Screening Station which allows automated image acquisition and data analysis of biological samples. Other terms for this field include image cytometry or automated microscopy.

The Scan^R is able to process large numbers of samples in an objective manner, enabling quantitative analysis across cells, wells, and samples. Although the total throughput is not as high as that of a flow cytometer it is still much higher than that of a typical microscope, allowing entire cell populations to be examined. Unlike flow cytometers, cells can be examined in situ, allowing morphological information from cells and tissue sections to be collected and used for analysis. However, the images acquired can be analyzed through the use of histograms, dot plots and even population gating as in flow cytometry.

Sample Types

Sample types which can be investigated with the Scan^R include both adherent and non-adherent cells and tissue sections. A climate-controlled chamber is available for living cells and time lapse experiments. 

Sample Templates

The Scan^R is very flexible with regard to what templates can be used for the samples. However, plates and dishes with optical bottoms are recommended for the best quality images and lowest background.

  • Microtiter plates (96- or 384-well)
  • Chambered coverslips
  • Standard slides
  • Petri dishes
  • Standard tissue culture plates

Excitation/Emission 

The scanR uses a SpectraX NIR LED Light Engine for fluorescence excitation. The possibility to use near infrared excitation/emission is unique amd makes the system particularly attractive for multiplexing fluorophores or for getting beyond regions of the spectrum where autofluorescence can be problematic. 

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Excitation Wavelengths

LED Color

Examples of Dyes or Proteins Which Could Be Used

395/25 nm

Violet

DAPI, Hoechst, Brillant Violets, other violet excitable dyes

438/29 nm

Blue

ECFP, Sytox Blue

475/28 nm

Cyan

EGFP, Alexa 488, FITC, CY2, (EYFP)

555/28

Green

Cy3, TRITC, Alexa 546/555, PE

*(mCh 575/25)

*(Orange)

*(mCherry, Alexa 694, Texas-Red)

635/22 nm

Red

Cy5, Alexa 633/647, APC, (Cy5.5), DRAQ5/7, SiR DNA

730/40 nm

Teal

NIR / CY7, Alexa 750, Zombie NIR

* Excitations for Cy3 and mCherry is an "either/or" choice and not simultanous.

   

 

Filter cubes, dichroics and emission filters are separate from the LED excitation system and can be used in different configurations.

Standard emission filters are listed below (others available upon request):

Emission Wavelengths

Examples of Fluorophores

433/24 nm

DAPI, Hoechst, BV421, Alexa 405

482/25 nm

eCFP, Sytox Blue

525/35 nm

eGFP, Alexa 488, FITC, CY2, PE

600/37 nm

Cy3, TRITC, Alexa 546/555

641/75 nm

mCherry, Alexa 594, Texas Red

680/42 nm

CY5, Alexa 633/647, APC, DRAQ5/7

809/81 nm

NIR / C7, Alexa 750, BV786

Additional filter cubes are also available (for example CY5.5, em 716/40 nm) but must be used in combination with LED excitation.

IHC

Although for quantitation fluorescence is always the first choice, it is also possible to examine/quantify standard histologically stained/IHC specimens (brightfield) using the scanR system in transmitted light mode.

If you have any questions about possible labelling strategies, we would be happy to help you.

 

Access

Only those investigators who have been previously trained to use the Scan^R Screening Station are allowed to use it without an operator. For information about reserving time on the instrument, contact one of us.

 

 

Further Information