Reagents

Our vast range of staining reagents and buffers can be used with the NucleoCounter® instruments. Some of their applications include disaggregation of cell clusters, dead/live labeling, and cell lysing. Click below to read more about each reagent’s properties and explore how they can be incorporated into your cell counting procedures.

How are our reagents used?

The reagents on our website are used for specialized protocols, such as counting aggregated cells or performing certain assays on our slide-based cell counters. Ensure to carefully read the application note accompanying each protocol to use our reagents as intended.

Our reagents are used for the following purposes:

  • Disaggregation: For highly aggregated cell types we recommend using the aggregated cell count protocol on one of our NucleoCounter® instruments. This involves using at least one of our buffers to disaggregate the cells without harming them.
  • Distinguishing live from dead cells: Fluorescent, nucleic acid-binding dyes, such as acridine orange (AO), propidium iodide (PI), or 4′,6-diamidino-2-phenylindole (DAPI) are used on our slide-based cell counters (NC-3000™ and the NC-250™). They provide accurate cell viability measurement and clear cell analysis.
  • Diameter estimation: AO readily permeates cells and binds to negatively charged cellular structures, which allows for estimation of cell diameter.
  • Neutralization buffers: to allow for optimal binding of fluorescence dyes, reagents are used to create the optimal chemical environment (e.g., Reagent B).
  • Whole blood analysis: when counting leukocytes in whole blood, buffy coats or bone marrow, our Blood Lysis buffer helps to lyse erythrocytes while staining the white blood cells.
  • Apoptosis assays: Some of our reagents are used in assays that provide information on cellular death, such as the annexin V assay, caspase assay, or mitochondrial potential assay.
  • Sperm analysis: For sperm cell counting using the SP-100™, Reagent S100 is used to ensure nuclei are stained prior to counting.
  • Yeats analysis: Reagent Y200 is used to prepare yeast cells for counting.

Note: We do not recommend using your own reagents for our cell counting procedures requiring reagent use.

How do the pH values of different reagents affect their functionality and usage?

The pH can greatly influence the intended function of a reagent.

The pH can affect a reagent’s reactivity, stability, and even solubility. Some of our reagents require a specific pH range for optimal performance. Examples include Solution 10, Reagent A100, Reagent B and Lysis 1 buffer.

We recommend following our protocols for optimal reagent use and cell counting and viability analysis.

Are there any specific storage conditions required for these reagents to maintain their efficacy?

Proper storage conditions are essential for consistent reagent performance. Each ChemoMetec reagent conveniently lists applicable storage conditions on the label of the product container itself, as well as on the material safety data sheet (MSDS). You can download a specific reagent’s MSDS from our web page; just type in the reagent part number in the search bar, click on right reagent to access its page and scroll to the link to the MSDS.

Note: Our reagents and consumables have expiration dates, which are stated on the product labels themselves alongside product lot numbers. Please do not use reagents and consumables past their stated expiration dates, in which case we cannot guarantee their efficacy.

What are the safety precautions to be taken while handling these reagents?

Prior to working with a ChemoMetec reagent, please review that reagent’s material safety data sheet (MSDS). The reagent’s MSDS includes the following important information:

  • Hazard description (if any) and relative composition of the reagent
  • How to safely handle and store the reagent
  • Reagent’s toxicological properties, what to do in case of exposure and how to administer first aid
  • How to safely dispose of the reagent

You can download a specific reagent’s MSDS from our web page; just type in the reagent’s part number in the search bar, access the specific reagent’s page and scroll to the MSDS link.

Please follow your institution’s guidelines regarding laboratory safety and good laboratory practices. Always consider the following:

  • Wear personal protective equipment (PPE): lab coats, gloves, and safety goggles.
  • Work in well-ventilated areas.
  • Verify reagent expiration date before use.
  • Initial and date reagent upon opening.
  • Store reagents as instructed by the MSDS and by reagent packaging.
How do these reagents enhance cell analysis?

Each ChemoMetec reagent serves a specific purpose. Some of them aid in obtaining an accurate and reproducible cell count, whereas others enable other types of cellular characterization.

Do specific solutions exist for detaching adherent cell lines and breaking apart cell clusters?

Yes. For an accurate cell count, highly aggregated cells must be de-clustered and in suspension. As part of their in-house processes, there are many different 3rd party reagents researchers use to detach adherent cells from flasks and bioreactor surfaces (beads, hollow fibers, strips, etc.).

Examples include, but are not limited to: accutase, dispase, collagenase and trypsin. Following dissociation, certain cell types may still exhibit clumping or clustering. Depending on the instrument, ChemoMetec offers specific protocols that address these situations to obtain a more accurate cell count. These protocols use specific reagents, which are stated in the protocol application notes.

Some examples are:

Instrument Protocol Context Reagent
NC-200™ and NC-3000™ Aggregated Cells Assay Aggregating cells Solution 10
NC-200™ and NC-3000™ A100 and B Heavily aggregating cells, cells grown in bioreactors Reagent A100, Reagent B
NC-200™ and NC-3000™ Blood Assay PBMCs in blood (does not count RBCs) Solution 17
NC-202™ Aggregated Cells Aggregating cells Lysis 1 buffer
NC-202™ Microcarriers Cells grown in bioreactors Lysis 1 buffer
NC-202™ Blood PBMCs in blood (does not count RBCs) Lysis 4
Which reagents are used in conjunction with others, and for what specific purposes?

ChemoMetec reagents are intended to be used with specific protocols, which are described in corresponding Application Notes. These can be accessed on our instruments’ software by clicking on the PDF icon in protocol selection menu. Some reagents work only in combination with others, or in specific contexts.

Some examples are:

  • Neutralizing and Stabilizing: The acidic nature of Reagent A100 requires neutralization with Reagent B for effective DNA quantification.
  • Tailored for Specific Cells: When quantifying cells in blood (e.g., apheresis samples), specific reagents are required for accurate cell counts on some ChemoMetec cell counters. For example, the NC-200™ requires Solution 17 and the NC-202™ requires Lysis 4 buffer. Likewise, reagents like Solution 10 and Lysis 1 buffer are intended for quantification of cells that tend to grow in clumps.