Single-Cell Analysis

Celselect Slides™ Enrichment Kit 2.0
User ManualIntroduction

Celselect Slides capture and isolate individual circulating tumor cells (CTCs) based on their size, utilizing
microfluidics and 140,000 individual microchambers. Isolated cells can be retrieved for downstream
analysis such as single-cell genomics, digital PCR (dPCR), or fluorescence in situ hybridization (FISH).
This protocol details use of Celselect Slides and reagents with the Genesis Cell Isolation System.
List of Provided Components and Consumables
Table 1 lists components and consumables included in theCelselect Slides Enrichment Kit 2.0.
Table 1. Celselect Slides Enrichment Kit 2.0 (catalog #17009455). Each kit
supports processing of four samples.
Box 1 of 2. Shipment and Storage Conditions: Room Temperature
Consumables

Quantity

Celselect Slides 2.0

4

Reagents cartridge piercer

4

Waste jars and tubing

4

Inlet funnel 2.04

Priming reservoir

4

Box 2 of 2. Shipment and Storage Conditions: 4°CFig. 1. The Genesis System.

Precautions■ Do not use reagents and consumables beyond the expiration
date on the label■ isually inspect each blood sample for clots before processing;V
discard any clotted samples■

Protect reagents from exposure to light sources■

Do not mix and match reagents from different lots/batches■

Reagents

Quantity

Dilution buffer

4

Reagents cartridge

4

Celselect Slides and Consumables (Box 1)

Each of the Celselect Slides and consumables is intended forsingle use.
Celselect Slides (Figure 2) use Bio-Rad’s proprietary CTC
capture and analysis technology. Each slide has one laminatedside and the other side contains inlet and outlet ports. Opticalcharacteristics closely mimic a glass slide, rendering the slide fitfor imaging.

 se protective barriers such as gloves, gowns, masks, and eyeU
protection when working with whole blood specimens

Fig. 2. Celselect Slides 2.0.

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List of Equipment Provided with the Genesis System

Blood and other reagents enter the Celselect Slides through theinlet funnel 2.0 (Figure 3).

Table 2. Equipment provided.

Fig. 3. Inlet funnel 2.0.

Equipment

Catalog Number

Quantity

Celselect Slides Station 2.012020055

2Reagents Cartridge Venter

CEL84093

1

Celselect Slides Imaging Adaptor 2.012019988

1

Wireless Barcode Scanner

CEL84092

1

Backflow Adaptor

12016521

2The Celselect Slides Station 2.0 (Figure 7) is a single piece with aspring-loaded clamp to keep the Celselect Slides in place and incontact with the thermal vapor chamber beneath.

The reagents cartridge piercer (Figure 4) is firmly placed in the drip
plate of the Genesis System to penetrate the bottom of the reagent
cartridge and allow reagents to enter the inlet funnel.

Fig. 7. Celselect Slides Station 2.0.
Fig. 4. Reagents cartridge piercer.

The waste jar (Figure 5) collects reagents and biohazardous waste.

The Celselect Slides Imaging Adaptor 2.0 (Figure 8) can hold up
to two Celselect Slides and is designed to fit into a standard PCR
plate format for use with an automated fluorescent microscope.

Fig. 5. Waste jar and tubing.

Fig. 8. Celselect Slides Imaging Adaptor 2.0.

The Genesis System priming reservoir (Figure 6) is used during
priming and again once the cells have been isolated.

The reagents cartridge venter (Figure 9) is designed to pierce thetop foil of the reagents cartridge for its use on the Genesis System.
Top-piercing is imperative to allow proper flow of reagents during
sample processing.

Fig. 9. Reagents cartridge venter.

The backflow adaptor (Figure 10) is designed to connect theGenesis System vacuum pump to the priming reservoir, producing
positive pressure to prime slides and retrieve captured cells during
CTC enrichment assays.

Fig. 6. Priming reservoir.

Celselect Slides Enrichment Reagents (Box 2)

The Genesis System dispenses reagents into the funnel in single
aliquots, and an entire set is provided for each run.
■■

Dilution buffer — the dilution buffer is used to retrieve target
cells after purification while maintaining the viability of theenriched sample
Reagents cartridge — the reagents cartridge is shipped
preloaded with all necessary reagents to facilitate cell capture
and purification© 2024 Bio-Rad Laboratories, Inc.

Fig. 10. Backflow adaptor.

2

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Celselect Slides Enrichment Kit 2.0 User ManualUser-Supplied Equipment, Consumables, and ReagentsThe equipment, consumables, and reagents listed below arenecessary to perform enrichment with Celselect Slides and canbe purchased from any vendor.
Equipment Supplied by User

Single-channel pipets (20, 200, and 1,000 μl)
Serological pipet controller■ Test tube rocker■ Test tube rack■
■Consumables and Reagents Supplied by User

Fig. 13. Place waste jar in position.

5 ml and 15 ml conical tubes■ Sterile disposable serological pipets (5 and 10 ml)■ Phosphate buffered saline (PBS), 1x, pH 7.4■ Pipet tips (20, 200, and 1,000 μl)■ 70% ethanol solution■4.

Attach the waste jar tubing to the waste jar (Figure 14).

Performing the Genesis System Self Test
1.

Power on the Genesis System using the switch found on theback bottom left corner of the instrument (Figure 11).

Fig. 14 . Attach waste jar tubing.

5.
Fig. 11. Genesis System back panel.

2.

Press tubing into the pinch valve until you feel it click into place(Figure 15).

Once the instrument is on, the Genesis System home screen(Figure 12) loads automatically and logs every run. From here,
the user can log in, gather system information, or shut down
the instrument.

Note: Log-in is not required.

Fig. 15. Press tubing into the pinch valve.

6.

Attach Genesis System tubing to the waste jar outlet(Figure 16).

Fig 12. Genesis System home screen.

3.

Place the waste jar in position at the base of the station bays
nearest the user (Figure 13).

© 2024 Bio-Rad Laboratories, Inc.

Fig. 16. Attach Genesis System tubing.

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7.

Press Start from the home screen to reach the Self Testscreen. Select each lane to initiate the Self Test (Figure 17).

Fig. 19. After a successful Self Test, the protocols can be selected.

11. Populate the data entry fields recording the reagents,
consumables, and slide identifiers using the barcode scanner(Figure 20).

Fig. 17. Run Self Test.

8.

A confirmation list will appear on the screen, which lists theactions/items the system looks for prior to running the Self
Test (Figure 18).

Fig. 20. Populate data for run.

Note: If you are unable to scan barcode/QR codes, use thefollowing to manually enter the lot numbers with identifiers:

Fig. 18. Confirm Run Self Test.

9.

a. Celselect Slides Enrichment Reagents (4-pack):“C” followed by the 6-digit lot number, e.g., C010101

Ensure that the configuration is correct, then click Next toinitiate the Self Test protocol.

b. Celselect Slides Enrichment Consumables (4-pack):“M” followed by the 6-digit lot number, e.g., M010101

10. When the Self Test is complete, select 2.0 Kit Enrichmentfrom the Select Protocol options (Figure 19).

c. Celselect Slides (4-pack): “S” followed by the 9-digit
serial number, e.g., S010101001
d. The Sample ID field is a free-form field allowing the user todefine a unique sample name for the run© 2024 Bio-Rad Laboratories, Inc.

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Celselect Slides Enrichment Kit 2.0 User ManualPriming and Assembly
Note: While the slide is priming, the reagents cartridge can bepulled from 4°C storage and warmed at 37°C for a minimum of
10 minutes in an incubator or bead bath.
1.

3.

Slide the reagents cartridge piercer until it clicks or snaps intoplace. Do this before placing the inlet funnel. The funnel is tootall to place the piercer after placing the funnel (Figure 23).

Place the slide into the slide station 2.0 by gently pullingthe spring-loaded tab to allow the slide to fall into place.
The notched end of the slide drops into the notched endof the slide station (Figure 21).

Fig. 23. Bottom piercer hitting inlet funnel if not added in correct order.

Fig. 21. Inserting the slide into the slide holder.

2.

4.

Tilt the inlet funnel under the piercer and attach it to the inlet
on the slide.

5.

Attach the backflow adaptor to the priming reservoir. Attach
the reservoir to the offset inlet on the right side of the slide.
Note: The backflow adaptor can be difficult to attach to thepriming reservoir when dry. Lightly wetting the O-ring or theinside of the priming reservoir with a gloved finger or lint-freewipe saturated with either water or 70% ethanol can help
the pieces slide together more easily. Using buffer is notrecommended.

Place the slide station containing the slide on theGenesis System. Firm pressure is required to snap the slide
station in place.
■■The slit in the station prevents incorrect placement he slide station assembly should be pushed all the way
Tback until its vertical arms (finger holds) meet the clamping
tabs (stoppers) on the Genesis System lane (Figure 22)

6.

Attach the Genesis System tubing to the backflow adaptor
tubing (Figure 24).

Fig. 22. Placing the slide station assembly into the Genesis System.

Fig. 24. Genesis System tubing attached to the backflow adaptor in
preparation for slide priming.

© 2024 Bio-Rad Laboratories, Inc.

5
Project Title: CelselectVendor: Troutt Visual Services
File Name: Fig_10_CelselectDate: 03/01/24Proof: 7 (final)

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7.

Sample Preparation and Processing
These steps can be done while the slide is priming. Slide priming
takes about 20 minutes to complete.

Pipet 8 ml 70% ethanol into the inlet funnel and start theenrichment run on the Genesis System.
Note: If 70% ethanol solution is prepared fresh, allow it todegas for at least 1 hour before use. Alternatively, the ethanol
solution can be purchased at 70% or made ahead of time.

8.

After priming, the instrument will prompt the user to remove
the backflow adaptor from the priming reservoir, discard
the ethanol, and replace the priming reservoir with the waste
jar tubing. Connect the luer of the waste jar tubing to the slide
completely by pressing down onto the outlet port. Twisting orover-tightening the luer may make it difficult to remove when
the run is complete.

If the biospecimen sample (blood) was kept chilled, allow thesample to acclimate to room temperature.

2.

Gently mix the biospecimen sample (blood) with a serologicalpipet several times. Draw 1–10 ml of sample per slide anddispense into appropriate conical tube.
Note: The sample will be diluted, so use a conical tube that
can accommodate your sample volume.

Note: The ethanol will be in the priming reservoir aftercompletion.
Best practice: To avoid introducing air into the slide, do notremove the ethanol that remains in the outlet port.
9.

1.

3.

Dilute the biospecimen sample 1:1 with room-temperature
1x PBS. For example, 7.5 ml blood + 7.5 ml 1x PBS.

4.

Gently mix the sample by pipetting up and down several times
with a serological pipet.

5.

Before dispensing the diluted sample into the funnel, gently
mix the sample by either inverting the tube several times orusing a serological pipet.

6.

Gently dispense the sample into the funnel and avoid
generating bubbles. If large bubbles occur, they can beremoved with a pipet.

7.

Important: Pierce the top of the reagent cartridge with thereagent cartridge venter before placing it onto the Genesis
System spindle.

Be sure to attach the Genesis System tubing to the waste jar,and press the waste jar tubing into the pinch valve (Figure 25).

Note: The cartridge has one lane labeled Start. This should
face the user.
8.

Gently rotate the cartridge until it slides down the spindle,
taking care not to manually twist the spindle itself.

9.

Press Resume on the Genesis System screen.

Cell Recovery
1.

Ensure that the dilution buffer is warmed for a minimumof 10 minutes at 37°C.

2.

When the run completes, the Genesis System will prompt
the user to remove the waste jar and reattach the priming
reservoir.

3.

Remove the waste jar tubing from the slide and place thepriming reservoir onto the inlet.

4.

Detach the Genesis System tubing from the waste jar.

5.

Pipet 4 ml of prewarmed dilution buffer into the primingreservoir.

6.

Attach the backflow adaptor to the priming reservoir andreattach to the Genesis System tubing.

7.

Press Resume on the Genesis System screen.

8.

The enriched sample can be retrieved by pipetting fromthe inlet funnel.

Fig. 25. Instrument loaded with slide, funnel, cartridge piercer, and waste
tubing properly routed through the pinch valve.

10. Pipet 8 ml 1x PBS into the funnel and press Resume on theGenesis System screen to wash the slide.
11. The instrument will now prompt the user to dispense thesample and place the reagents cartridge.

© 2024 Bio-Rad Laboratories, Inc.

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Cleaning the Genesis System
Clean the Genesis System after each use by wiping the instrument
deck with either 10% bleach solution or 70% ethanol on atowelette.
Cleaning the Slide Stations
Clean the slide stations after each use by wiping them with either
10% bleach solution or 70% ethanol on a towelette.
Disposal of Kit Consumables and Reagents
All used kit consumables and reagents should be treatedas biohazardous waste and disposed of in accordance withappropriate internal laboratory safety and environmental guidelines.
Specimen Collection, Transport, and Storage
Collect blood aseptically into EDTA tubes.
Caution: Once the tube is opened, process blood immediately.
Table 3. Specimen collection, transport, and storage.
Enumeration
ProtocolTransportation
Temperature

Storage
Temperature

EDTA tubes

Ship on ice toincrease the stabilityof the cells

Refrigerate sample
and bring to room
temperature, 15–30°C(59–86°F), before
processing

Shelf Life
Process within
24 hours aftercollection

BIO-RAD and CELSELECT SLIDES are trademarks of Bio-Rad Laboratories, Inc. incertain jurisdictions. All trademarks used herein are the property of their respective
owner. © 2024 Bio-Rad Laboratories, Inc.
For research use only. Not for use in diagnostic procedures.

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