3um Flow Red-Fluo Microspheres RCY3UM-630 0.5 %

3µm Flow Red-Fluo Microspheres for Flow Cytometry

SHBC RCY3UM-630 Flow Red-Fluo Microspheres are 3µm far-red fluorescent microspheres developed for red-laser flow cytometry research, fluorescence channel evaluation, instrument method development, bead-based assay research, and bulk reagent production.

The microspheres are designed for excitation at approximately 630nm and provide a broad far-red emission range of 665–720nm. This spectral profile is suitable for flow cytometers equipped with compatible red lasers and far-red fluorescence detectors.

RCY3UM-630 is supplied as a 0.5% solids suspension and is available for laboratory evaluation, pilot-scale development, repeated production, and bulk supply.

Quick Product Answer

RCY3UM-630 is a 3µm far-red fluorescent microsphere suspension with 0.5% solids, approximately 630nm excitation, and 665–720nm emission. It is intended for flow cytometry research, red-laser fluorescence detection, instrument-setting studies, bead-based method development, and research reagent manufacturing.

Product Highlights

• Product name: Flow Red-Fluo Microspheres

• Catalog number: RCY3UM-630

• Nominal particle diameter: 3µm

• Excitation wavelength: 630nm

• Emission wavelength: 665–720nm

• Fluorescence region: Far red

• Solids content: 0.5%

• Primary application: Flow cytometry research

• Supply capability: Sample, pilot, and bulk production

• Intended use: Research use only

RCY3UM-630 Product Overview

RCY3UM-630 consists of 3µm fluorescent microspheres that generate a far-red fluorescence signal under compatible red-laser excitation.

The 3µm particle diameter provides a clearly defined microsphere population for many conventional flow cytometry platforms. Compared with submicron particles, 3µm microspheres generally produce stronger optical scatter and may be easier to separate from electronic noise and particulate background.

This makes RCY3UM-630 suitable for research teams developing or evaluating:

• Red-laser fluorescence detection

• Far-red detector settings

• Scatter and fluorescence gating

• Flow cytometer method setup

• Fluorescent particle tracking

• Bead-based assay platforms

• Particle concentration optimization

• Reagent development workflows

• Instrument performance comparison

• Batch-to-batch fluorescence evaluation

The product can be used by biotechnology companies, flow cytometry laboratories, instrument developers, assay manufacturers, universities, research institutes, and diagnostic reagent development organizations.

RCY3UM-630 is supplied for research and development. Any use as a formal calibration material, absolute counting standard, quantitative fluorescence standard, or regulated diagnostic control requires additional qualification and validation.

Technical Specifications

Additional lot-specific information may be available according to the confirmed production specification.

Project documentation may include, where applicable:

• Certificate of Analysis

• Particle-size test results

• Fluorescence test results

• Solids-content information

• Product handling instructions

• Safety Data Sheet

• Storage recommendations

• Lot and production information

Particle composition, surface chemistry, preservative system, particle-size coefficient of variation, fluorescence intensity limits, packaging configuration, and shelf life should be confirmed using the quotation, technical data sheet, or lot-specific Certificate of Analysis.

Far-Red Fluorescence with 630nm Excitation

RCY3UM-630 microspheres are designed for excitation near 630nm and emit fluorescence across the 665–720nm wavelength range.

This excitation and emission profile is suitable for red-laser flow cytometry systems using a compatible far-red detector.

Potentially compatible red-laser wavelengths may include:

• 630nm

• 633nm

• 635nm

• 637nm

• 640nm

The fluorescence response obtained at each laser wavelength may differ. Compatibility should therefore be verified using the intended flow cytometer.

The broad 665–720nm emission region may overlap several far-red detector configurations. Depending on the instrument, suitable optical filters may include filters centered within approximately:

• 660–680nm

• 665–695nm

• 670–710nm

• 680–720nm

• 690–735nm

The actual signal depends on:

• Laser wavelength

• Laser power

• Detector type

• Detector gain or voltage

• Bandpass filter

• Dichroic mirror configuration

• Particle concentration

• Sample flow rate

• Buffer background

• Instrument optical alignment

Researchers should review the actual optical configuration rather than relying only on channel names such as APC, FL4, far red, red fluorescence, or R-channel.

Why Choose 3µm Fluorescent Microspheres?

Easier Detection on Conventional Flow Cytometers

The 3µm particle diameter is considerably larger than nanoparticles and submicron beads.

On many conventional flow cytometers, 3µm particles may generate stronger forward-scatter and side-scatter signals than 500nm particles. This can make the microsphere population easier to locate and gate during initial instrument setup.

Instrument performance still varies, and detection should be confirmed on the intended platform.

Suitable for Red-Laser Channel Evaluation

RCY3UM-630 provides a far-red fluorescence signal under red-laser excitation.

The product may be evaluated when researchers need to confirm that a red laser and the associated far-red detector can identify a consistent fluorescent particle population.

Useful for Method Development

A defined fluorescent microsphere population can help researchers establish:

• Detector gain settings

• Fluorescence thresholds

• Scatter gates

• Sample flow rates

• Particle dilution levels

• Event-rate limits

• Background exclusion rules

• Analysis templates

Suitable for Bead-Based Research

The 3µm size range is commonly considered for bead-based flow cytometry research because the particles can be detected individually while providing a physical surface that may support future assay-development strategies.

When biomolecule coupling is required, an appropriate functional surface must be selected and validated.

Appropriate for Bulk Reagent Development

RCY3UM-630 can be evaluated in early-stage research and then transferred into pilot or bulk production after the required technical specifications have been confirmed.

Key Product Features

3µm Particle Diameter

The nominal 3µm size provides a detectable microsphere population for flow cytometry method development, fluorescence analysis, and particle-based research.

Far-Red Fluorescence

The microspheres are designed for approximately 630nm excitation and 665–720nm emission.

Red-Laser Compatibility

RCY3UM-630 is intended for evaluation on flow cytometers equipped with compatible red lasers.

0.5% Solids Content

The supplied concentration allows researchers to prepare working dilutions according to the required event rate and application.

Broad Far-Red Emission

The broad emission range provides flexibility when selecting detectors and filters that overlap the far-red spectrum.

Research-to-Production Supply

The product can support initial sample testing, laboratory optimization, pilot production, and bulk purchasing.

Custom Project Support

Alternative particle sizes, concentrations, fluorescence intensities, surface chemistries, and packaging formats may be discussed for qualified development projects.

Flow Cytometry Applications

Red-Laser Fluorescence Detection

RCY3UM-630 may be used to evaluate whether a flow cytometer’s red laser and far-red detector can detect a defined fluorescent particle population.

Researchers may compare:

• Fluorescence intensity

• Detector response

• Signal-to-background separation

• Event distribution

• Histogram width

• Day-to-day measurement stability

Instrument Setup Research

The microspheres may support the development of consistent instrument setup procedures.

Possible research uses include:

• Locating the particle population

• Adjusting forward-scatter gain

• Adjusting side-scatter gain

• Setting far-red detector voltage

• Establishing a fluorescence threshold

• Creating a standard analysis gate

• Checking sample flow stability

• Comparing low, medium, and high flow rates

RCY3UM-630 should not automatically be described as a certified instrument-calibration standard unless the supplied product has been specifically tested and documented for that purpose.

Far-Red Channel Verification

The microspheres can be used to determine whether the selected detector filter overlaps the product’s 665–720nm emission range.

The user may evaluate:

• Detector sensitivity

• Filter compatibility

• Background separation

• Signal saturation

• Acquisition repeatability

• Channel-to-channel spillover

Formal compensation controls normally require controls that match the fluorophore and labeling characteristics of the assay. RCY3UM-630 should therefore be validated before being used as a compensation control.

Fluorescence Trigger Development

In some research workflows, fluorescence may be used as an acquisition trigger instead of scatter.

RCY3UM-630 can be evaluated for:

• Fluorescence-triggered acquisition

• Trigger-threshold optimization

• Background discrimination

• Low-fluorescence event exclusion

• Comparison of scatter and fluorescence triggering

Particle Dilution and Event-Rate Studies

The 0.5% suspension can be diluted to prepare a range of particle concentrations.

A dilution series may help identify conditions that provide:

• Stable event rates

• Minimal coincidence

• Clear particle resolution

• Reproducible fluorescence

• Acceptable acquisition time

• Low sample consumption

Fluidic-System Evaluation

Fluorescent microspheres may be evaluated when studying sample introduction and particle movement through a flow cytometer or microfluidic system.

Potential observations include:

• Event-rate stability

• Sample carryover

• Flow interruption

• Particle recovery

• Tube-to-tube consistency

• Fluidic cleaning effectiveness

Bead-Based Assay Development

When supplied with or developed using an appropriate surface chemistry, 3µm fluorescent microspheres may support bead-based assay research.

Potential assay-development directions include:

• Antibody immobilization

• Antigen immobilization

• Protein-binding studies

• Nucleic acid capture research

• Ligand-receptor interaction studies

• Multiplex particle coding

• Fluorescent carrier particle development

The standard RCY3UM-630 surface chemistry must be confirmed before planning a coupling procedure.

Fluorescence Microscopy and Particle Tracking

In addition to flow cytometry, the microspheres may be evaluated in compatible fluorescence microscopy, imaging, microfluidic, filtration, and particle-tracking studies.

Application suitability depends on the microscope excitation source, filter set, detector sensitivity, sample matrix, and required particle behavior.

Compatible Lasers and Detection Channels

Recommended Excitation

A red laser close to the specified 630nm excitation wavelength is recommended.

Potential laser lines include:

RCY3UM-630 is not primarily designed for excitation with 405nm, 488nm, or 561nm lasers unless separate spectral testing confirms sufficient excitation.

Recommended Detection Region

Select a detector whose optical filter overlaps the 665–720nm emission range.

A filter centered near 680nm, 690nm, or 700nm may be suitable when its bandwidth captures a sufficient part of the emission spectrum.

The best detector cannot be selected using the center wavelength alone. Both the center wavelength and filter width should be considered.

For example:

• A 670/30 filter collects approximately 655–685nm.

• A 695/40 filter collects approximately 675–715nm.

• A 710/50 filter collects approximately 685–735nm.

These are examples of filter ranges rather than guaranteed instrument recommendations.

APC-Like Detection Channels

The product’s far-red emission may overlap detector channels commonly associated with APC-like fluorescence.

However, RCY3UM-630 should not be described as chemically identical or spectrally identical to APC. Compatibility should be based on the actual excitation and emission profile rather than the fluorophore channel name.

Suggested Flow Cytometry Workflow

The following workflow is a general starting point. Instrument settings and sample preparation should be optimized for the intended application.

1. Prepare the Flow Cytometer

Clean and prime the instrument according to the manufacturer’s instructions.

Confirm that the fluidic system is stable and that the red laser and far-red detector are available.

2. Run a Buffer Blank

Use the same buffer that will be used to dilute the microspheres.

Record the blank event rate in:

• Forward scatter

• Side scatter

• Far-red fluorescence

• Any other monitored channels

A buffer blank helps identify particulate contamination, electronic noise, buffer fluorescence, and carryover from previous samples.

3. Resuspend the Microspheres

Before sampling, gently mix the vial until the suspension is homogeneous.

Suitable initial methods may include:

• Gentle inversion

• Controlled vortexing

• Brief mild bath sonication when necessary

Avoid excessive foaming, overheating, prolonged sonication, or harsh mechanical treatment.

Do not rely on visual appearance alone to confirm complete dispersion.

4. Prepare a Dilution Series

Prepare multiple working dilutions in clean tubes.

For example, a preliminary study may include several tenfold or stepwise dilutions.

The exact dilution depends on:

• Original solids concentration

• Flow cytometer sample rate

• Desired event rate

• Acquisition time

• Application

• Background level

Start with a low concentration and gradually increase it when necessary.

5. Use a Low Sample Flow Rate

A low flow rate is recommended during initial optimization because it may improve population resolution and reduce coincident events.

After suitable conditions are established, medium or high flow rates can be evaluated.

6. Locate the Particle Population

Begin with forward-scatter, side-scatter, and far-red fluorescence plots.

Adjust the scatter and fluorescence settings gradually until the particle population is clearly separated from the buffer background.

7. Select the Far-Red Detector

Choose a detector filter that overlaps the 665–720nm emission range.

Increase detector voltage or gain carefully to avoid saturating the signal.

8. Establish a Gate

Create a gate around the main microsphere population.

Exclude:

• Buffer noise

• Debris

• Possible aggregates

• Very high-intensity coincident events

• Electronic artifacts

9. Evaluate Coincidence

Run at least two or three particle dilutions.

If the apparent fluorescence intensity, scatter position, or event distribution changes significantly as the sample is diluted, coincidence may be present.

Continue dilution until the measurement becomes concentration independent within the required experimental range.

10. Record All Settings

For reproducibility, record:

• Instrument model

• Laser wavelength

• Detector filter

• Detector voltage or gain

• Scatter settings

• Trigger channel

• Trigger threshold

• Sample flow rate

• Dilution factor

• Acquisition time

• Number of collected events

• Buffer composition

Using RCY3UM-630 in Bead-Based Assay Development

RCY3UM-630 may be considered as a fluorescent particle platform during bead-based flow cytometry research.

Before assay development, confirm whether the product has a plain, carboxyl, amino, streptavidin, epoxy, or other functional surface.

Biomolecule Immobilization

If an appropriate surface chemistry is available, the microspheres may be evaluated for immobilizing:

• Antibodies

• Antigens

• Proteins

• Peptides

• Enzymes

• Oligonucleotides

• Nucleic acid probes

• Other ligands

The coupling method must be selected according to the surface functional group and the biomolecule.

Assay Development Factors

Important parameters may include:

• Coupling buffer

• Buffer pH

• Biomolecule concentration

• Activation-reagent concentration

• Coupling time

• Blocking agent

• Washing procedure

• Storage buffer

• Microsphere concentration

• Target sample matrix

• Non-specific adsorption

• Fluorescence retention

Multiplex Flow Cytometry Research

Different microsphere populations may potentially be distinguished using:

• Particle diameter

• Fluorescence color

• Fluorescence intensity

• Surface chemistry

• Internal coding strategy

For a multiplex assay, each population should remain clearly separated after coupling, blocking, storage, and sample incubation.

Reporter fluorescence should also be separated from the RCY3UM-630 far-red identification signal.

RCY3UM-630 Compared with 500nm Fluorescent Microspheres

Both 3µm and 500nm microspheres can be used in flow cytometry research, but they are suitable for different experimental priorities.

Choose RCY3UM-630 When:

• A clearly detectable far-red particle population is required.

• The project uses a conventional flow cytometer.

• The research focuses on red-laser detector setup.

• A micron-sized bead platform is preferred.

• Bead-based assay development is being considered.

• Scatter and fluorescence gating are both required.

Choose 500nm Microspheres When:

• The project specifically focuses on submicron particles.

• Small-particle detection performance is being studied.

• The instrument has suitable nanoparticle or extracellular-vesicle detection capability.

• Fluorescence-triggered acquisition is a major research objective.

Quality Control and Batch Consistency

For reagent-development and bulk-production projects, the required quality specifications should be agreed upon before manufacturing.

Potential quality-control parameters may include:

• Nominal particle diameter

• Particle-size distribution

• Particle-size coefficient of variation

• Suspension appearance

• Solids content

• Dispersion performance

• Aggregate level

• Excitation spectrum

• Emission spectrum

• Relative fluorescence intensity

• Fluorescence distribution

• Flow cytometry histogram

• Lot-to-lot fluorescence comparison

• Microbial control

• Storage stability

• Accelerated stability

• Packaging integrity

Not every parameter is included in every standard product specification. Required limits should be discussed during technical evaluation.

Flow Cytometry Evaluation

A flow cytometry QC procedure may evaluate:

• Main population position

• Fluorescence median or mean

• Distribution width

• Percentage of gated particles

• Aggregate population

• Background separation

• Run-to-run repeatability

Instrument settings should remain unchanged when comparing different production lots.

Calibration and Traceability

RCY3UM-630 should not automatically be represented as:

• A NIST-traceable particle-size standard

• A certified fluorescence-intensity standard

• A MESF-assigned standard

• An absolute particle-count standard

• A regulated diagnostic control

• A certified flow cytometer calibration bead

These claims require specific testing, value assignment, documentation, and traceability.

Project-specific qualification may be discussed when a formal reference-material application is required.

Bulk Supply and Custom Manufacturing

SHBC supports fluorescent microsphere projects from laboratory evaluation through pilot production and bulk manufacturing.

Research Samples

Samples may be used to evaluate:

• Red-laser compatibility

• Far-red detector compatibility

• Fluorescence intensity

• Particle dispersion

• Flow cytometry gating

• Sample dilution

• Assay feasibility

Pilot Production

Pilot batches may support:

• Method optimization

• Stability research

• Packaging evaluation

• Initial lot comparison

• Assay transfer

• Customer verification

Bulk Production

After the technical specification is confirmed, bulk manufacturing can be arranged according to the customer’s expected demand.

Information required for bulk evaluation may include:

• Required quantity

• Annual forecast

• Target package size

• Delivery schedule

• Application

• Flow cytometer configuration

• Required fluorescence level

• Surface chemistry

• Buffer requirements

• QC specifications

• Documentation requirements

Custom Manufacturing Options

Depending on technical feasibility, customization may include:

• Alternative particle diameters

• Different excitation wavelengths

• Different emission wavelengths

• Adjusted fluorescence intensity

• Alternative solids contents

• Plain microsphere surfaces

• Carboxyl-functionalized surfaces

• Amino-functionalized surfaces

• Other functional surfaces

• Customer-specific packaging

• Customer-specific QC criteria

• OEM labeling

• ODM product development

Final feasibility depends on the complete technical requirement and order volume.

Handling and Storage Recommendations

For best performance, follow the product-specific label, technical data sheet, and Certificate of Analysis.

General recommendations include:

• Store protected from light.

• Refrigerated storage at 2–8°C is generally recommended unless otherwise specified.

• Do not freeze unless freeze-thaw stability has been confirmed.

• Keep the vial tightly closed.

• Store the container upright.

• Mix the suspension before use.

• Use clean pipette tips and sample tubes.

• Avoid contaminating the original container.

• Minimize prolonged exposure to strong light.

• Avoid repeated unnecessary temperature changes.

• Do not allow the microspheres to dry.

• Do not return diluted material to the original container.

Resuspension

If settling occurs during storage:

1. Allow the vial to reach the recommended handling temperature.

2. Mix gently by inversion.

3. Apply controlled vortexing when necessary.

4. Inspect the suspension for visible aggregates.

5. Evaluate a small sample by flow cytometry before beginning the full experiment.

Brief mild bath sonication may be evaluated if ordinary mixing does not provide sufficient dispersion.

Light Protection

Because the product is fluorescent, unnecessary exposure to strong laboratory light should be minimized.

Working dilutions should be prepared shortly before use when possible.

Freezing

Freezing may cause irreversible particle aggregation or changes in suspension performance.

Do not freeze the product unless the supplied documentation specifically permits frozen storage.

Frequently Asked Questions

What is RCY3UM-630?

RCY3UM-630 is a 3µm far-red fluorescent microsphere suspension developed for flow cytometry research. It has approximately 630nm excitation, 665–720nm emission, and 0.5% solids.

What is the particle diameter?

The nominal particle diameter is 3µm.

Lot-specific particle-size information should be confirmed using the corresponding technical document or Certificate of Analysis.

Which laser should be used?

A red laser near 630nm is recommended.

Flow cytometers equipped with 633, 635, 637, or 640nm lasers may also be suitable, but compatibility should be confirmed experimentally.

Which fluorescence channel should be used?

Use a far-red detector whose optical filter overlaps the 665–720nm emission range.

Channel names vary by instrument, so the actual filter wavelength should be checked.

Can it be detected in an APC channel?

The emission range may overlap some APC-associated detection channels.

However, RCY3UM-630 is not necessarily spectrally identical to APC. Compatibility should be determined from the instrument’s laser and filter configuration.

Can a 488nm laser excite RCY3UM-630?

The product is specified for approximately 630nm excitation.

It should not be assumed to provide sufficient fluorescence under 488nm excitation unless separate spectral testing confirms this behavior.

Is RCY3UM-630 suitable for conventional flow cytometers?

The 3µm diameter is suitable for evaluation on many conventional flow cytometers.

Actual detection depends on the instrument optics, laser, detectors, filters, fluidics, and settings.

Can it be used for instrument alignment?

It may be evaluated during instrument-setting and comparative alignment research.

It should not be sold or described as a certified alignment standard unless appropriate qualification, calibration, and documentation are provided.

Is it a fluorescence calibration bead?

Not automatically.

A certified fluorescence standard normally requires assigned fluorescence values, defined uncertainty, traceability, and supporting documentation.

RCY3UM-630 is supplied primarily as a research fluorescent microsphere unless otherwise specified.

Can it be used for absolute counting?

Absolute counting requires a verified particle number concentration and an appropriate counting protocol.

The 0.5% solids value alone is not sufficient to establish an absolute number of particles per milliliter. Do not use the product as an absolute counting standard unless a validated particle-count value has been supplied.

Can it be used for compensation?

The product may be evaluated for fluorescence-channel research, but formal compensation controls should closely match the assay fluorophore and staining characteristics.

The user should validate RCY3UM-630 before using it as a compensation control.

Can antibodies be coupled to the microspheres?

Antibody coupling requires a suitable functional surface.

Please confirm whether the required product should have carboxyl, amino, streptavidin, epoxy, or another surface chemistry.

Can it be used in multiplex assays?

Yes, it may be evaluated as one fluorescent particle population in multiplex flow cytometry research.

The particle population must remain distinguishable from other bead populations and reporter fluorescence.

Why choose 3µm instead of 500nm particles?

The 3µm particles are generally easier to resolve by scatter on conventional flow cytometers and are suitable for bead-based assay and instrument-setting research.

The 500nm particles are more suitable for submicron detection and small-particle method development.

What solids concentration is supplied?

RCY3UM-630 is supplied at 0.5% solids.

The appropriate working concentration should be determined by dilution according to the required event rate.

How should the product be mixed?

Mix gently by inversion or controlled vortexing before sampling.

Brief mild bath sonication may be evaluated if necessary. Avoid excessive foaming and overheating.

How should it be stored?

Store protected from light and follow the storage instructions on the product documentation.

Refrigerated storage is generally recommended, and freezing should be avoided unless otherwise specified.

Is bulk production available?

Yes. RCY3UM-630 can be supplied for sample evaluation, pilot development, repeat orders, and bulk production.

Can the concentration or packaging be customized?

Alternative solids concentrations and packaging formats may be discussed according to technical feasibility, order quantity, and project requirements.

What information is needed for a quotation?

Please provide:

• Catalog number RCY3UM-630

• Required quantity

• Expected annual demand

• Preferred package size

• Flow cytometer model

• Red-laser wavelength

• Far-red detector filter

• Intended application

• Required surface chemistry

• Required fluorescence intensity

• QC requirements

• Documentation requirements

• Delivery destination

Request a Sample or Bulk Quotation

RCY3UM-630 3µm Flow Red-Fluo Microspheres provide a far-red fluorescent particle solution for red-laser flow cytometry research, instrument-setting studies, fluorescence channel evaluation, bead-based assay development, and research reagent manufacturing.

The product combines:

• 3µm nominal particle diameter

• Approximately 630nm excitation

• 665–720nm far-red emission

• 0.5% solids

• Flow cytometry research compatibility

• Sample and pilot supply

• Bulk manufacturing capability

• Custom-development support

For technical evaluation or quotation, provide the intended flow cytometer configuration, application, required quantity, packaging requirements, surface chemistry, and quality-control specifications.

Product Name: 3µm Flow Red-Fluo Microspheres
Catalog Number: RCY3UM-630
Particle Diameter: 3µm
Excitation Wavelength: 630nm
Emission Wavelength: 665–720nm
Solids Content: 0.5%
Primary Application: Flow Cytometry Research
Supply Capability: Samples, Pilot Batches, and Bulk Production
Intended Use: Research Use Only. Not for diagnostic or therapeutic use.