Systems And Methods For Improved Imaging And Fluorescence In Flow Cytometry And Other Applications

An imaging system, comprising:

• a sample region configured to receive a sample;
• a laser illumination train configured to provide laser illumination to the sample region to give rise to fluorescence of a sample in the sample region;
• an optical illumination train configured to provide imaging illumination to illuminate a sample in the sample region for imaging;
• a fluorescence train configured to collect fluorescence evolved in the sample region;
• a sample region lens in optical communication with the sample region,
• the sample region lens being configured (i) to direct illumination collected from the sample region to the fluorescence train and (ii) to direct illumination from the optical illumination train to the sample region;
• an image capture train comprising an imager configured to collect an image of a sample in the sample region illuminated by the imaging illumination.

An imaging system, comprising:

• a sample region,
• the sample region having a flow cell therein, the flow cell configured to communicate a sample therein;
• a sample region lens, the sample region lens defining a field of view of a designated portion of the flow cell;
• a plurality of laser pinholes, the plurality of laser pinholes arranged along a direction of flow within the flow cell and being configured to illuminate a sample transiting the designated portion of the flow cell;
• an illumination source, the illumination source configured to illuminate the sample at an image position within the designated portion of the flow cell.

A method, comprising:

• illuminating a sample in a designated region of a flow cell with laser illumination from one or more laser pinholes operating at different wavelengths;
• illuminating the sample in the designated region of the flow cell with imaging illumination from an optical illumination train configured to provide imaging illumination to illuminate the sample for imaging;
• with a lens, (i) directing, to a fluorescence train, illumination emitted from the sample in response to the excitation illumination, and (ii) directing, to the sample region, imaging illumination from the optical illumination train;
• and
• collecting, with an image capture train, an image of the sample;
• and
• collecting, with the fluorescence train, illumination emitted from the sample in response to the excitation illumination,
• wherein, relative to the sample, the laser illumination is along a first direction, the imaging illumination is along a second direction, the collecting the image of the sample is along a third direction, and collecting the illumination emitted from the sample in response to the excitation illumination is in a fourth direction.