LUTHOR single cell dispenser
LUTHOR single cell dispenser is the ideal companion for all LUTHOR HD Single-cell RNA sequencing workflows. This compact instrument fits on any laboratory bench. Easy to install and to operate, it ensures that cells are gently dispensed, one at a time.
Complete workflow
Conveniently integrate your full workflow solution from cell isolation to RNA-Seq library
Low cell input
You can start dispensing with as low as 100 input cells in your suspension
Compact size
Fits on any bench or even under the cell culture hood
Keep track of your cells
Each well is easily visualized for presence of a single cell
LUTHOR single cell dispenser efficiently isolates a full plate of individual cells in only 10 minutes by dispensing them as gently as manual pipetting.
The core technology of the LUTHOR single cell dispenser is based on impedance measurement of cells passing through the proprietary dispensing tip which records the electrical signal. This allows to monitor that only one cell is dispensed per well and directly identifies doublets and debris which can be excluded from further analysis.
A dedicated, intuitive, software is provided free of charge with the instrument to ensure seamless operation and streamlined analysis.
Instrument Specifications and Compatibilities
LUTHOR single cell dispensing Kit
The LUTHOR single cell dispensing Kit (Cat. No. 231.04) contains reagents and dispensing tips sufficient for four experiments. The consumables kit is required for use of the instrument.
Compatible RNA-Seq library preparation kits for NGS experiments from single-cells
Performance
The LUTHOR single cell dispenser associates ease of use with high reliability for optimal sampling of cells. Subsequent LUTHOR HD library preparation provides a unique workflow for high definition scRNA-Seq with unprecedented sensitivity.
Figure 2 | Comparative analysis between fresh and fixed DU145 cells.
Upper panel: Gene detection rates shown as average number of genes detected at increasing read depth. 8 single DU145 cells were analyzed from dispensing experiments using suspensions of A) fresh and B) fixed cells. Libraries for high-definition scRNA-Seq were generated with the LUTHOR HD Library Prep Kit and sequenced on Element’s AVITI platform. Gene detection was monitored at a threshold of ≥ 1 CPM (Counts Per Million). Read depth is expressed as raw reads. Standard deviation is indicated in light green.
Lower panel: Representative correlation analyses (orthogonal regression of gene-based collapsed and mapped reads) obtained from one fresh cell and one fixed cell. The R² value was computed from C) 28 (fresh vs. fresh), D) 28 (fixed vs. fixed) or E) 64 combinations (fresh vs. fixed) of single cells (sample size: 8 cells per condition).
Figure 3 | Comparison of read statistics from HEK293T cells isolated with the LUTHOR single cell dispenser and by flow cytometry. A) Percentage of total mapped reads and reads on genes. B) Percentage of reads mapping uniquely, multi-mapping and unmapped reads. C) Percentage of exonic*, intronic*, and intergenic reads*. D) Percentage of protein coding**, mitochondrial ribosomal RNA**, and cytoplasmic ribosomal RNA*. Cell input was 10,000 cells (at 20 cells/ml) for LUTHOR single cell dispenser and 1 million cells (at 1,000 cells/ ml) for the flow cytometer (BD FACSMelody, with 100-mm nozzle and 34,000 drops/sec). scRNA-Seq libraries were prepared with the LUTHOR HD Library Prep Kit. 6 samples per isolation method were analyzed at 1 M reads / sample.
*based on mapped reads; **based on exonic reads.
Figure 4 | Comparative analysis between HEK293T cells isolated with the LUTHOR single cell dispenser and by flow cytometry.
Upper panel: Gene detection rates shown as average number of genes detected at increasing read depth. 6 single HEK293T cells isolated with A) LUTHOR single cell dispenser and B) a flow cytometer. Libraries for high-definition scRNA-Seq were generated with the LUTHOR HD Library Prep Kit and sequenced on Illumina’s NextSeq2000 platform. Gene detection was monitored at a threshold of ≥ 1 CPM (Counts Per Million). Read depth is expressed as raw reads. Standard deviation is indicated in light green.
Lower panel: Representative correlation analyses (orthogonal regression of gene-based collapsed and mapped reads) obtained from single HEK293T cells isolated with either the LUTHOR single cell dispenser or a flow cytometer. The R2 value was computed from C) 15 (LUTHOR dispenser replicates), D) 15 (flow cytometer replicates) or E) 36 combinations (LUTHOR dispenser vs. flow cytometer) of single cells (sample size: 6 cells per condition). Cell input was 10,000 cells (at 20 cells/ml) for LUTHOR single cell dispenser and and 1 million cells (at 1,000 cells/ ml) for the flow cytometer (BD FACSMelody, with 100-mm nozzle and 34,000 drops/sec).
Workflow
Step 1: Prepare cells
Eukaryotic cells (down to 100 cells / experiment)
Step 2: Filter and transfer into scd Buffer
Cells are filtered to remove debris, then transferred into scd Buffer (dispensing buffer)
Step 2: Filter and transfer into scd Buffer
Cells are mixed with the scd Buffer, then the LUTHOR single cell dispenser aspirates them through the scd Tip.
Step 4: Dispense cells
Cells are dispensed, one by one, into each well of a 96-well plate (containing LUTHOR lysis buffer). Impedance change generates an electrical signal each time a cell passes through the scd Tip aperture.
Step 5: Analyze
Each well is easily visualized for presence of a single cell.