An interview with Daniel Schieffer, Expertise Director, DeNovix
Why are cells at the moment counted with slides? What number of instances are the slides usually used and what occurs to them after they’re used?
Till lately, cells had been usually counted on a reusable quartz hemocytometer. With the appearance of automated cell counters 12-15 years in the past, disposable slides grew to become widespread. This was primarily a approach for these cell counters to load and depend slides. Whereas they’re handy for automated programs, they introduce an extra ongoing value for labs, and the single-use plastic additionally has a big environmental footprint.
Picture credit score: Shutterstock | Watiporn Yenchum
Every slide is designed to solely be used as soon as. There could be two counts per slide, however they’re all single-use and are routinely disposed of after use. We have heard tales of individuals attempting to reuse single-use slides; nevertheless, for the sake of accuracy this isn’t really helpful by slide producers. It additionally provides considerably to the hands-on time required to depend cells.
How does the CellDrop remove disposable slides? The place are the cells saved and the way is full cleansing achieved between runs?
To enhance this method DeNovix mixed one of the best of the outdated and new, changing plastic slides with two everlasting, optical grade sapphire surfaces. The surfaces are positioned parallel to at least one one other by a excessive precision screw to kind a chamber analogous to a standard hemocytometer. The core expertise is tailored from our DS-11 Sequence of microvolume spectrophotometers and customers are instantly accustomed to the load, measure and wipe clear workflow.
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Researchers dispense the cells into the chamber, press depend and the evaluation is carried out routinely. Afterwards, the pattern is solely wiped away with a dry laboratory wipe and the CellDrop is prepared for the following pattern. One of many nice options of the CellDrop is a big, high-definition display screen with a stay preview of the measurement chamber. This enables the person to immediately affirm that the surfaces are clear.
Picture credit score: DeNovix
How does the efficiency examine to slide-based cell counters?
The slide-based counters use plastic slides that introduce inconsistencies between counts. For instance, if a researcher has 10 plastic slides, there will likely be 10 slight variations between every of the slides. The CellDrop system makes use of a stable sapphire floor that’s constant every time it’s used, and the chamber heights are routinely calibrated each time the instrument is turned on.
It is very reproducible run to run. As well as, cell counts on CellDrop are very quick in comparison with different automated counters, so there’s an actual efficiency improve within the person expertise.
Picture credit score: DeNovix
What functions does the CellDrop work with?
Our important functions are at the moment in cell viability, i.e. stay/useless counts for mammalian, yeast and bug cell tradition. There are two fashions of the CellDrop out there—twin fluorescence and brightfield, or brightfield solely.
For brightfield viability measurements, scientists use a dye comparable to Trypan blue to stain cells. Trypan blue has its limitations, so for labs requiring better specificity and accuracy in addition to a better vary of functions, we suggest using fluorescence. The CellDrop can use frequent fluorophores comparable to Acridine orange and Propidium iodide (AO/PI), the place the fluorescent AO stains stay cells inexperienced and PI stains useless cells pink.
Utilizing this utterly takes the guesswork out of counting stay/useless cells or counting main cells in samples which can be doubtlessly contaminated. For instance, it avoids counting pink blood cells or different bits and particles {that a} typical Trypan algorithm or handbook counting may confuse for a cell, which results in an inaccurate depend.
You can even analyze GFP tranfected cells with CellDrop, offering the p.c viability and transfection effectivity. The platform is designed to be open to a variety of functions throughout cell biology but additionally environmental, meals and brewing industries.
Can the CellDrop be used with hazardous and infectious samples?
There are two methods to deal with hazardous or infectious samples with the CellDrop. First, we designed it to take a seat in a biosafety cupboard. Researchers with bio-safety degree 2 or bio-safety degree 3 lab can put their CellDrop straight into the hood the place they’re coping with hazardous cells.
Analysis continues as regular, utilizing a typical cleansing protocol on the CellDrop after the cell samples have been counted. For instance, if 10% bleach is used to sterilize the gear and space used, this may be sprayed on a paper towel to wipe down the CellDrop after use.
We additionally enable for using disposable slides throughout the CellDrop for the aim of infectious or hazardous materials. If there’s a pattern that for no matter motive a lab actually would not wish to contact the CellDrop straight, a spread of slides from totally different producers can be utilized.
On this case, the researcher would put the slide on high of the reusable floor after which deal with the cells on the slide, depend it, after which throw it into the hazardous waste with out the pattern ever having touched the CellDrop.
What software program and algorithms/AI include the CellDrop cell counter? How do these assist scientists?
DeNovix software program is designed by life scientists for all times scientists. We work arduous to make the software program not solely intuitive to make use of, but additionally very highly effective. Researchers can shortly depend and analyze outcomes instantly on board the instrument. The counting functions and the entire embedded algorithms are preinstalled on CellDrop counters, so there is no such thing as a want for added tablets, computer systems or cloud-based software program.
Inside every counting software, we’ve included a default protocol that works properly for most typical cell strains. Clients can simply optimize settings for various cell varieties after which save these in protocols for added counts. Outcomes are saved onboard for straightforward retrieval for re-analysis or audit functions.
What limitations are there for the CellDrop system?
The system was engineered as a cell counter. So like several optically designed instrument, there are decision constraints because of the physics of the system. We will reliably depend cells down 4 microns and there actually aren’t any limitations in comparison with what you are able to do with a handbook depend or a unique automated cell counter.
We have designed it to offer one of the best of each worlds—the pace of automation however with out the extra value and inconvenience of disposable slides. In the event you want single-use disposable slides for any motive the CellDrop can accommodate that, nevertheless it additionally provides researchers the flexibleness to make use of a reusable system for almost all of analysis they run.
What does the longer term maintain for DeNovix Cell Counters?
We are going to proceed to broaden the functions and performance of the CellDrop with our free software program updates. We work intently with our clients and have already got some nice concepts that we’re planning to implement.
Extra broadly, DeNovix goes to proceed to search for locations the place we will innovate to avoid wasting clients’ money and time and cut back ache factors within the lab. We wish to simplify instrumentation and supply instruments scientists want to have however that aren’t cost-effective given the present expertise or want to purchase into a brand new consumable stream.
The place can readers discover extra info?
https://www.denovix.com/celldrop/
About Daniel Schieffer
Dan joined NanoDrop Applied sciences Inc. in 2006 whereas finishing his BS in Biotechnology at College of Delaware. After contributing to the event of the NanoDrop 8000 and NanoDrop 2000 Dan accepted a instructing and analysis assistantship at Villanova College.
At Villanova he helped develop a novel circulation cytometry-based FRET system to elucidate the function of cell membrane group in CD4+ T Cell activation.
Dan then moved right into a analysis assistantship function on the College of Pennsylvania the place he carried out excessive throughput genetic screens to find novel immune system pathways that regulate pathogenesis in insect born viruses earlier than becoming a member of DeNovix in 2013. He has an MS in Immunology from Villanova College.
