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

General Specifications
General Specifications
Power SupplyExternal, 12 V, 11.5 A, 138W
Plate holder2 plates (96 or 384 wells)
96-well plate
384-well plate (customized setup)*
Minimal cell number100 cells**
CalibrationNot required
CleaningNot required
Sample prep. (time to dispense)10 min (96-well plate)
40 min (384-well plate)
Cell parametersSize, doublets
Input voltage (External power supply)100 - 240 V~, 50 - 60 Hz
Input current (External power supply)max 2.0 A
Input voltage (Instrument)12.0 V
Max. power (Instrument)40 W
Well plates compatibility96-well plate: non-skirted, semi-skirted (with plate holder)
96-well plate: fully skirted (without plate holder)
384-well plate: fully skirted (without plate holder)
Any 96-well and 384-well plate that meets ANSI/SLAS 1-2004 through ANSI/SLAS 4-2004 standards is compatible.
Loading tubeSarstedt 0.5 ml, 150 ml
Loading volumeper 30s load. time @1.5x10⁴ cells/mL: 10µL, 200 cells
Dispensing volume (receiving well)Down to 5 µl
Particle range detection capabilityDielectric, from 8 µm to 25 µm
*contact: support@lexogen.com
** LUTHOR single cell dispenser can be adapted down to 100 cells as initial input. Preset programs are readily available, and if needed, Lexogen offers additional support to customize new programs tailored to each experimental setup.
Compressed Air Specifications
Compressed Air Specifications
Minimum operating pressureLess than 0.2 psi
Environmental Conditions
Environmental Conditions
Temperature5 °C to 40 °C (41 °F to 104 °F)
Maximum relative humidity80% for temperature up to 31 °C
Instrument Measurements
Instrument Measurements
Instrument weight11.1 kg
Instrument Size328 x 271.5 x 369 mm
Additional Hardware
Additional Hardware
Computer screen with HDMI cableRequired, not included
KeyboardRequired, not included
MouseRequired, not included
Consumables
Consumables
LUTHOR single cell dispensing KitCat. No. 231.04, sufficient for 4 dispensing experiments

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.

Consistent Dispensing Accuracy – Even with Initial Low Cell Numbers
The LUTHOR single cell dispenser gently dispenses cells into individual wells, even when your cell input is very low (down to 100 cells). The standard protocol for starting suspensions with higher cell numbers achieves a plate filling rate of 91 % of wells receiving one single cell (Fig. 1 A), whereas a plate filling rate of 86 % is achieved with the low-volume protocol which allows processing of suspensions with low initial cell numbers (Fig. 1 B).
Report of cell count per well following DU145 cell dispensing using the LUTHOR single cell dispenser with either standard protocol (A) or low-volume protocol for limited cell input – down to 100 cells (B) achieving 91 % and 86 % monoclonality, respectively.
Figure 1 | Typical output of cell preparations dispensed into 96-well plates with the LUTHOR single cell dispenser. A) Human DU145 cells were dispensed with the standard protocol (3,000 cells in 150 μl scd Buffer, i.e., 20 cells / μl). B) DU145 cells dispensed using the low-volume protocol (400 in 40 μl scd Buffer, i.e., 10 cells / μl). Wells labelled in green with “1” contain a single cell. Wells labelled in red with “2C”, “3C”, “4C” respectively contain two, three, or four cells.
Fresh or Fixed: Consistent Library Quality
The LUTHOR single cell dispenser is also compatible with fixed cells. Dispensing and scRNA-Seq with LUTHOR HD provide highly consistent and comparable results whether fresh or fixed cell inputs are used (Fig. 2).
Fresh and fixed cells show comparative gene detection rates over various read depths and high replicate correlation between two individual fresh cells, two individual fixed cells as well as across conditions (fresh vs. fixed) following dispensing with the LUTHOR single cell dispenser, scRNA-Seq library preparation with LUTHOR High-Definition Single-Cell 3’ mRNA-Seq and sequencing on Element’s AVITI platform.
Fresh and fixed cells show comparative gene detection rates over various read depths and high replicate correlation between two individual fresh cells, two individual fixed cells as well as across conditions (fresh vs. fixed) following dispensing with the LUTHOR single cell dispenser, scRNA-Seq library preparation with LUTHOR High-Definition Single-Cell 3’ mRNA-Seq and sequencing on Element’s AVITI platform.

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).

Excellent Alternative to Flow Cytometry-assisted Cell Dispensing
The LUTHOR single cell dispenser provides a bench-top alternative to flow cytometers for dispensing of individual cells within minutes. Single-cell RNA-Seq data sets from individual cells obtained using the LUTHOR single cell dispenser or a flow cytometer show highly consistent and similar read statistics (Fig. 3).
Read statistics assessing mapped reads, reads mapped to genes, mapping classes (multi-mapped, uniquely mapped, unmapped reads), feature classes (exonic, intronic and intergenic reads), as well as protein-coding, rRNA and mitochondrial rRNA reads obtained from LUTHOR High-Definition Single-Cell 3’ mRNA-Seq libraries using cells isolated either with a flow cytometer or with the LUTHOR single cell dispenser. Results are highly comparable and of equally high quality regardless of the cell isolation method used.

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.

Unsurpassed Gene Detection Sensitivity and Precision with Both Flow Cytometry-assisted and LUTHOR single cell dispenser-assisted Cell Isolation
Cells isolated with the LUTHOR single cell dispenser and prepped with the LUTHOR HD 3’ mRNA-Seq kit provide the same high-definition sensitivity RNA-Seq data as cells isolated with a flow cytometry sorter (Fig. 4).
Cells isolated with the LUTHOR single cell dispenser, or a flow cytometer show comparative gene detection rates over various read depths regardless of the isolation method. In addition, high replicate correlation is observed between two individual cells isolated using the LUTHOR single-cell dispenser, two individual cells using flow cytometry as well as across conditions (dispenser vs. flow cytometer) following scRNA-Seq library preparation with LUTHOR High-Definition Single-Cell 3’ mRNA-Seq and sequencing on Illumina NextSeq2000.
Cells isolated with the LUTHOR single cell dispenser, or a flow cytometer show comparative gene detection rates over various read depths regardless of the isolation method. In addition, high replicate correlation is observed between two individual cells isolated using the LUTHOR single-cell dispenser, two individual cells using flow cytometry as well as across conditions (dispenser vs. flow cytometer) following scRNA-Seq library preparation with LUTHOR High-Definition Single-Cell 3’ mRNA-Seq and sequencing on Illumina NextSeq2000.

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.

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