Easy banking of blood (and other sample types) for subsequent analysis by CyTOF is a core competency of our Smart Tubes and other products. No RBC lysis or centrifugation are required at the site banking the samples.
One of the best ways to see how Smart Tube products are used is to check out the publications that have used the system to bank their samples. We've started compiling a list of these studies. In fact, our team is especially excited about CyTOF so nearly all of the studies featured on the following page used CyTOF to analyze their Smart Tube banked samples:
http://www.smarttubeinc.com/publications.htm
The first one listed is "A Streamlined CyTOF Workflow To Facilitate Standardized Multi-Site Immune Profiling of COVID-19 Patients" which details an exciting new way to stain and bank COVID-19 samples in the Smart Tube Proteomic Stabilizer PROT1
Next is "Landscape of coordinated immune responses to H1N1 challenge in humans" where volunteers were infected with H1N1 to better understand the immune response to this virus.
Fourth down is a study by Janssen Pharma where samples from the clinical trial of Daratumumab to treat multiple myeloma were banked in Smart Tubes.
You can navigate to each article by clicking on the thumbnail image at the far left. Tables of the antibodies used to analyze the banked samples by CyTOF can be downloaded by clicking the PDF icon to access the PDF version of the table or by clicking on the XLS icon to download a copy of the Excel spreadsheet version.
You might also want to check out the following published protocols that detail use of Smart Tube products:
Protocol published by Janssen Pharma: Smets, Tina, et al. "Deep Profiling of the Immune System of Multiple Myeloma Patients Using Cytometry by Time-of-Flight (CyTOF)." Multiple Myeloma. Humana Press, New York, NY, 2018. 47-54.
Behbehani, Gregory K. "Cell Cycle Analysis by Mass Cytometry." Cellular Quiescence. Humana Press, New York, NY, 2018. 105-124.
Fernandez, Rosemary, and Holden Maecker. "Cytokine-Stimulated Phosphoflow of Whole Blood Using CyTOF Mass Cytometry." Bio-protocol 5.11 (2015).
While these protocols are focused on mass cytometry, there are very few steps that are specific to mass cytometry (note the similarity to our official protocols which are for conventional flow cytometry)
To cover all the bases, below is a detailed summary of Smart Tube products which discusses the multiple options for banking samples (whole human blood; bone marrow; cell suspensions; murine blood; murine tissue samples) as well as a paragraph on how Smart Tubes operate as well as links to some of the published studies that banked their samples in Smart Tube products.
There are multiple options for banking samples with Smart Tube products:
Option 1: Using Smart Tubes with the compact Base Station automation instrument
Option 2: Using the Smart Tubes manually, without the Base Station
Option 3: Using a bottle of the same Proteomic Stabilizer (name: PROT1) which is employed in the Smart Tubes
Option 4: Using our other reagent system, the Stable-Lyse Stable-Store reagent system to bank samples
Summary:
Each Smart Tube contains a sealed ampoule of our Proteomic Stabilizer, PROT1. At the site collecting the samples the user will simply place blood in each Smart Tube, put the Smart Tubes into the Base Station automation instrument, and hit the "Start" button to initiate the pre-programmed protocol. The Base Station presses on the Smart Tubes to break the ampoule inside and release the Proteomic Stabilizer into the blood sample and the Base Station rotates the tubes to ensure good mixing with the Proteomic Stabilizer. The Base Station only requires 10 minutes to complete the Stabilization reaction whereupon the Smart Tubes can be removed from the Base Station and directly transferred to -80C for storage with out further handling. In the event that a user is not available to transfer the samples to -80C, the Base Station holds the samples at 4 degrees until the user transfers the tubes to -80C. The Base Station can execute the protocol without user intervention so the entire sample prep process takes no more tech time than is required to load blood into the tubes making this system appropriate for busy facilities where time limited and studies where temporal control and protocol adherence are critical.
Downstream Processing Details: The samples, frozen and stored in Smart Tubes, are thawed and incubated in a buffer that lyses the erythrocytes (erythrocytes are not lysed before the samples are frozen). The resulting leukocytes can be handled as if they were formaldehyde fixed, although surface antibody staining has been reported to be superior to conventional fixation methods. The system is compatible with alcohol based and detergent based permeabilization methods.
A growing number of clinical studies have used Smart Tubes to bank their clinical samples (blood and bone marrow). One of these studies collected patient samples from over 12 different hospitals (30 Base Stations total). This is one of a growing number of studies that had us make their reagents of interest into instantly dissolving LyoSpheres and load them into Smart Tubes so that they could incubate the live patient blood samples with these reagents for a period of time before the Smart Tubes released the PROT1 proteomic stabilizer into the sample thereby preserving the sample for later analysis.
In addition to the proteomic stabilizer "PROT1" which was used for the studies above, we also have a second reagent system for banking whole blood, the "Stable-Lyse Stable-Store" reagent system. The following is a comparison of the PROT1 reagent system and the Stable-Lyse Stable-Store reagent system:
1. PROT1 System:
a. A single liquid reagent is added to the blood sample allowing it to be banked at -80C. No centrifugation or water bath is required to bank samples.
b. This reagent system is available in bottle format or in our "Smart Tubes" which provides the option to use our Base Station Automation Instrument to standardize sample handling and optionally, allow controlled incubation of the live blood samples with reagents that induce biological states of interest (ie. antigens, small molecule drugs, cytokines, cross-linking antibodies). Note that we can make biologics including cytokines or antibody cocktails (even cocktails of conjugated antibodies) or small-molecule drugs into lyophilized pellets known as "LyoSpheres" that instantly dissolve in blood; we can load these LyoSpheres into the Smart Tubes and seal them in foil pouches.
c. Has been on the market since 2009
d. Several published clinical studies have used Smart Tubes to bank their clinical samples (blood and bone marrow). I've included references to these papers at the bottom of this email. One of these studies collected patient samples from over 12 different hospitals (30 Base Stations total). This is one of a growing number of studies that had us make their reagents of interest into LyoSpheres and load them into Smart Tubes so that they could incubate the live patient blood samples with these reagents for a period of time before the Smart Tubes released the PROT1 proteomic stabilizer into the sample thereby preserving the sample for later analysis.
e. Smart Tubes can be operated manually, without the Base Station automation instrument. The blood sample would be placed into the Smart Tube and then the cap screwed on securely. After an optional incubation at 37C (ie. to use drugs or biologics to induce biological states in the blood sample) the user will hold the Smart Tube between thumb and forefinger and flex it to break the ampoule inside the Smart Tube and release the Proteomic Stabilizer into the blood sample. The Smart Tube is then incubated at room temperature for 10 minutes and then transferred directly to -80C for frozen storage.
Alternatively, the proteomic stabilizer PROT1 (purchased in bottle format) can be pipetted into a blood sample that has previously been placed in conventional plasticware (ie. a FACS tube or conical tube).
2. Stable-Lyse Stable-Store System:
a. Two different liquid reagents are added in series to the blood sample allowing it to be stored for days at 4C and banked at -80C. Some users have successfully left samples prepped with our Stable-Lyse Stable-Store system at 4C for over 20 days before freezing them at -80C. Some users are now reporting that they are also getting acceptable results with test samples stored at room temperature (have even tested storage at 37C) for days or even weeks --- you would have to test this to ensure that you achieve acceptable results. Just like with the PROT1 System, no centrifugation or water bath is required to bank samples. This ability to leave the sample at 4C for extended periods of time is a key advantage of the Stable-Lyse Stable-Store System. May give slightly better staining performance than the PROT1 system for some surface markers and for phospho-STAT antibodies.
b. Available in bottle format. Not available in Smart Tubes. A fluidics cartridge based system that automates the use of the Stable-Lyse Stable-Store reagent system is under development.
Stable-Lyse V2 in bottle format; 250ml Size; use 350ul per 250ul of whole blood then add 1000ul of Stable-Store (catalog number STBLYSE2-250)
Stable-Store V2 in bottle format; 250ml Size; use 1000ul per 250ul of whole blood after adding 350ul of Stable-Lyse V2 (catalog number STBLSTORE2-250)
Stable-Store V2 in bottle format; 1000ml Size; use 1000ul per 250ul of whole blood after adding 350ul of Stable-Lyse V2 (catalog number STBLSTORE2-1000)
You can view our publications page on our website : https://www.smarttubeinc.com/publications.htm
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