Introducing Laminar Wash Technology for Improved Flow Cytometry
Cell processing technology enabling superior consistency and robust flow cytometry data using centrifuge-less washing.
The Laminar Wash technology provides an advantage to scientists in processing suspension cells for flow cytometry. Eliminating the usage of the centrifuge has tremendously improved workflows and time management prior to FACs analysis. In the conventional flow cytometry protocol which utilizes the centrifugation method, immune cells undergo numerous washes that generate significant stress to cells. This in turn increases the alteration of biomarker pathways as well as cell loss.
With the Laminar Wash technique, cells undergo a gentle wash without added stress, allowing a higher retention of cells for acquisition and cell sorting. In addition, cell surface markers are not modified and unbound antibodies are washed off efficiently, producing cleaner and better segregated data. For high throughput flow cytometry labs, they will be able to achieve consistency by reducing variation from manual handling.
The Laminar Wash system has been successfully demonstrated on various applications including immunophenotyping and CyTOF. Some of the lymphocytes and cell surface markers tested, with improved cell retention and data are as follows:
• PBMCs
• Antigen presenting cells (CD40) such as B-cells (CD34, CD45) and dendritic cells
• T-cells – Cytotoxic T cells, Helper T cells (CD3, CD4, CD8)
• Monocytes
• Macrophages
• Cell lines: HEK293T, Yeast cerevisea, CHO, U937, Ramos, Molt4, THP-1, MOM13
• Primary Mouse Tumor Cells
• Mouse Primary Splenocytes
• Colorectal Tumor cells
• Circulating Tumor cells
• Isolated Nucleii
• Thymic Epithelial cells from Mouse
• Mouse Brain cells
• Nucleated red blood cells from maternal blood / cord blood
See How Laminar Wash Technology Works:
See Laminar Wash Technology in Action:
Here we see a plate dispensed with two drops of green fluorescent polystyrene beads and two drops of food coloring solution. After 20 min of rest, beads all settle to the bottom of wells on the plate. Then a user places the plate in the Laminar Wash HT1000 System and hits the start button. The plate goes in and the fluidics head comes down to a plate. In this run, the case of a washer was lifted intentionally to show the movement of the head and plate inside. The head has 192 nozzles total, 2 nozzles per well, one for dispensing and one for aspiration. During 2 minute duration, it performs 7 times of dispensing and aspiration each. When the wash is complete, a plate comes out. In the pictures, you can see that the food coloring solution completely gone while green fluorescent beads are retained on the plate. You may wonder how come the ink was washed away while beads were retained. Just to clarify, the beads are not attached to a plate. They are naturally and gently sitting on a plate by gravity without any adhesion
Direct HTS acquisition by flow cytometer on Laminar Wash 96-well Plates
Equivalent acquisition efficiency and stability as U-bottom plate
Efficient cell mixing even with low cell concentration
Mouse splenocytes (250,000 cells) seeded per well of Laminar Wash plate with accessory grid or U-bottom plate, and volume topped up, brought to 150uL with FACS buffer. Cells were acquired by HTS on BD Celesta flow analyzer, and gated for lymphocytes by FSC-SSC scattering. (Acquisition speed: 1.0uL/s. Mixing volume: 100uL. Mixing speed: 100uL/s. No. of mixes: 5
Placement of grid increases volume per well for efficient cell mixing by HTS probe
Washing Whole Blood Without a Centrifuge – Human and Mouse
Lysing and washing red blood cells using traditional microtiter plates or tubes may result in significant cell loss and damage. There is now a better way to lyse RBCs while achieving clean antibody staining and lysis as good as or better than traditional methods. The new Laminar Wash system offers centrifuge-less blood lysis and leukocyte washing on a single platform, with efficient RBC and debris removal. Whole blood lysis performed by Laminar Wash systems has similar leukocyte retention compared to lysis with centrifugation, whether enumerated by CD45 staining or FSC-SSC scattering
The scatter plot above is derived from human whole blood lysed and stained on Laminar Wash System vs. Microtiter plate vs. FACS tube (L to R).
The scatter plot above is derived from mouse blood with TER119 vs. CD45 using Laminar Wash HT1000 system vs. Microtiter plate vs. FACS tube (L to R). Note the clean staining using Laminar Wash System.
Laminar Wash Technology lets you achieve:
- Consistency by reducing variation from manual handling
- Time savings by completing cell washing in 3 minutes
- Reduced cell stress due to no centrifugation
- Easy integration into your lab automation