Nexcelom Bioscience

978-327-5340

Measure % RFP Positive Cells in Samples Containing GFP, RFP and Non-Expressing Cells

A 96-well plate containing samples with varied % of RFP, GFP and non-expressing HeLa cells were counted using Celigo. Bright field, green fluorescence and red fluorescence channels were used.

Step 1. Acquire & View Images on Celigo Image Cytometer

View whole-well cell images

fluorescent co culture assay View whole well cell images

Double click any well on the well map to display whole-well images. Zoom in to see details as shown below in cell image examples.


Bright field image of well D6, zoomed view

bright field images for co culture assay

RFP fluorescent image of well D6, zoomed view

step 2 RFP fluorescent co culture assay

Cell image examples from wells with varied RFP+ cell populations

GFP+, RFP+

GFP+, RFP+ cell image

RFP+

RFP+ cell image

Counted RFP+

Counted cells in RFP+ image

GFP+, RFP+

GFP+, RFP+ cell image

RFP+

RFP+ cell image

Counted RFP+

Counted cells in RFP+ image

GFP-, RFP+

GFP-, RFP+ cell image

RFP+

RFP+ cell image

Counted RFP+

Counted cells in RFP+ image

Step 2. Gate and Circle Counted Cells Automatically

Celigo gating and segmentation interface

Celigo gating and segmentation interface

Overlay of bright field, GFP+ and RFP+ cells identified by Celigo image processing software. GFP vs RFP intensity scatter plot is also shown. When the gate on the scatter plot is modified by dragging, the counted cell image is updated automatically.


Example Plots

GFP vs. RFP Scatter Plot

GFP vs. RFP Scatter Plot

RFP Intensity Histogram

RFP Intensity Histogram


GFP vs. RFP Scatter Plot

GFP vs. RFP Scatter Plot

RFP Intensity Histogram

RFP Intensity Histogram


GFP vs. RFP Scatter Plot

GFP vs. RFP Scatter Plot

RFP Intensity Histogram

RFP Intensity Histogram

Step 3. Review Plate Level Results for Each of the Wells in the 96-Well Plate

Results are displayed in a plate-based format

Plate Level Results for 96-Well Plate

Step 4. Export Results for Each of the Wells in the 96-Well Plate and Produce Data Plot

Results exported in Excel

Exported results for 96-well plate

Percent Expression of Fluorescent Proteins

Percent Expression of Fluorescent Proteins
Top