anvajo: Thank you very much for taking the time to do our interview. Could you please introduce yourself?
Dr. Shvartsman: My name is Dmitry Shvartsman. I am Chief Technology Officer at Cellaria. We work in the field of cancer research, patient testing and treatment development by creating in-vitro disease models which represent the specific nature of the biology of a patient. Our models originate from tissue from a patient to provide details that are of clinical interest and help to define the characterization of the model. In the case of cancer, for example, the models possess features, both molecular and phenotypic, which closely resemble the patient’s. Similarly, for RNA-mediated derivation of iPS cell lines, our models permit the study of disease-specific pathways of activity. All in all, we want to characterize disease models to determine therapies that more effectively suit the needs of each patient.
anvajo: That’s very interesting! How many employees do you have?
Dr. Shvartsman: There are currently 11 employees at the company.
anvajo: And how did you hear about anvajo?
Dr. Shvartsman: I got in touch with the company via LinkedIn. Since we work with different cell types and perform cell counting, it made sense for us to test the fluidlab R-300.
anvajo: That’s nice. For which cells did you use the fluidlab R-300 and which functions of the device did you use?
Dr. Shvartsman: 9 different cell lines and 4 cell types were used. The cells we counted with the fluidlab were fibroblasts, cancer cells, immune T cells and iPSCs, originating either from human, chicken, or pig. We received a fluidlab R-300 with acella 50 slides and acella 100 slides for 10 and 20 µL load volumes respectively and 3 adapters for slides and cuvettes. For cell count experiments we used acella 50 slides and to test the reproducibility of the device, we used acella 50 and acella 100 slides.
anvajo: So, you counted the cells with the fluidlab. How did you proceed and what were your results?
Dr. Shvartsman: Yes. We wanted to evaluate the intra slide reproducibility and the inter slide reproducibility. For the intra slide reproducibility we loaded acella 50 and 100 slides with cancer cells. Each slide we read 6 times, always laying them on the flat side between readings. Manually we loaded a hemocytometer with cancer cells and read one large square 6 times. To test the inter slide reproducibility we loaded 6 separate acella 50 and 100 with cancer cells and read each of them once. We also loaded a hemocytometer with cancer cells 6 times and read one large square with each load. We were very content with the results, showing CVs between 1% and 5% for the intra slide reproducibility and 7% - 10% for the inter slide reproducibility which is very much.
anvajo: That’s nice to hear! Were there any other criteria you used to evaluate the instrument?
Dr. Shvartsman: Yes, other criteria were the ease of setup and operation, and we also compared the counts of the fluidlab R-300 with manual counts to evaluate the accuracy by using acella 50 slides.
anvajo: How did it perform concerning the criteria?
Dr. Shvartsman: Regarding the cell count it was in line with the manual counts at concentrations above 50 000 cells/mL. When the concentration was below this value, we got mixed results (Fig.1). But we were still happy with it, since it was accurate for cell counts routinely observed at our company. Apart from that, the setup and use of the cell counter was very easy. We were also able to download the software datalab and to connect the fluidlab to WiFi.
anvajo: I’m glad it worked so good for you. Were there also challenges you had to face with the device?
Dr. Shvartsman: Yes, we work a lot with activated T cells. For proliferation we activate T-cells with microspheres and wanted to use the fluidlab to distinctively count cells and beads. This was challenging for the device.
anvajo: Yes, we can imagine! I assume that the cells and beads have a similar size, right? How did this affect the count?
Dr. Shvartsman: Exactly. The hydrogel, also microsphere, is linked to antibodies for activation. Manually counting the cells in the presence of these microspheres is easy as the cells and spheres look quite different. But we wanted to know if the fluidlab R-300 can identify the cells and microspheres as distinct populations. To determine this, a T cell titration was prepared with and without the beads and counts were determined manually using the acella 50 slide. The results showed that with no microspheres, the device does a good job in counting cells except at the 1:128 dilution where manually only 1 cell per field is observed. The instrument gives a nice warning that the count may not be correct.
anvajo: Very interesting! So, what is your overall conclusion about working with the fluidlab?
Dr. Shvartsman: For us it worked very well. It was easy to handle, and we were satisfied with the reproducibility as well as with the accuracy of the cell count. For T cells, at a high cell to microbead ratio, the instrument did well with and without beads. As this ratio decreases, of course it becomes very challenging for every automated system. But it would be fantastic if we could use the instrument for this application.
anvajo: Are you planning any other experiments with the fluidlab R-300?
Dr. Shvartsman: Yes, will continue to collect more data. We are also trying to implement some sort of low density seeding up to single cell seeding and we will see what the threshold of the machine is in terms of total counts, to not only take a high pellet, but also a very low volume to see if we can read it.
anvajo: We are curious how the results will turn out! Thank you very much for the insights. We wish you all the best an keep us updated!