500nm Flow Red-Fluo Microspheres RCY500NM-630 0.5%

500nm Flow Red-Fluo Microspheres for Flow Cytometry

SHBC RCY500NM-630 Flow Red-Fluo Microspheres are 500nm far-red fluorescent polymer microspheres developed for flow cytometry research, submicron particle analysis, fluorescence detection method development, and flow-based assay development.

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

RCY500NM-630 is supplied as a 0.5% solids suspension and is available for research evaluation, pilot-scale development, and bulk production.

Quick Product Summary

• Product name: Flow Red-Fluo Microspheres

• Catalog number: RCY500NM-630

• Nominal particle diameter: 500nm

• Excitation wavelength: 630nm

• Emission range: 665–720nm

• Solids content: 0.5%

• Fluorescence region: Far red

• Primary application: Flow cytometry research

• Supply capability: Samples, pilot batches, and bulk production

• Intended use: Research use only

RCY500NM-630 Product Overview

RCY500NM-630 consists of fluorescent polymer microspheres with a nominal diameter of 500nm. The fluorescent material is incorporated into the particles to provide a detectable far-red signal when excited by a compatible red laser.

The combination of a submicron particle diameter and far-red fluorescence makes these microspheres suitable for laboratories developing flow cytometry methods for small-particle detection.

Compared with cell-sized flow cytometry beads, 500nm particles can be used to evaluate instrument sensitivity, fluorescence-triggered detection, threshold settings, background control, sample dilution, and gating strategies for submicron particle populations.

The broad 665–720nm emission range also gives researchers flexibility when selecting detector filters that overlap the far-red fluorescence spectrum.

RCY500NM-630 is intended for research organizations, flow cytometry laboratories, instrument developers, reagent manufacturers, biotechnology companies, and assay development teams requiring consistent fluorescent microsphere supply.

Technical Specifications

Lot-specific information, available surface chemistry, packaging options, particle-size characterization, fluorescence testing, and supporting documentation can be discussed according to project requirements.

Far-Red Fluorescence with 630nm Excitation

RCY500NM-630 microspheres are designed for excitation around 630nm and emit fluorescence across the 665–720nm range.

This excitation and emission profile is suitable for flow cytometers equipped with red laser lines such as 630, 633, 635, 637, or 640nm, provided that the instrument detector and optical filter overlap the emission spectrum.

The far-red fluorescence region offers several practical benefits for flow cytometry research:

• Compatibility with commonly available red-laser flow cytometers

• Separation from many green and yellow fluorescence channels

• Potential integration into multicolor experimental designs

• Reduced interference from shorter-wavelength fluorescent reporters

• Flexible detector selection across the 665–720nm emission region

• Suitable fluorescence triggering for submicron particle analysis

Actual fluorescence intensity and channel compatibility should be confirmed on the intended flow cytometer because laser power, optical configuration, detector sensitivity, filter bandwidth, threshold settings, and sample concentration can affect the final signal.

Why Use 500nm Fluorescent Microspheres in Flow Cytometry?

Submicron Particle Method Development

The 500nm diameter provides a relevant particle size for developing and optimizing flow cytometry methods focused on submicron particles.

These microspheres may be used when evaluating instrument settings for research involving extracellular vesicle models, synthetic nanoparticles, microvesicle-sized particles, drug-delivery particles, viruses, liposomes, and other small fluorescent objects.

They are not intended to represent the biological or chemical properties of these samples, but they can provide a consistent synthetic particle population for method development.

Fluorescence-Based Detection

Submicron particles may produce relatively weak forward-scatter and side-scatter signals on conventional flow cytometers. Fluorescent labeling can provide an additional detection parameter and may support fluorescence-triggered acquisition.

RCY500NM-630 microspheres can therefore be used to study the relationship between particle fluorescence, optical scatter, electronic threshold settings, background events, and instrument sensitivity.

Red-Laser Compatibility

The 630nm excitation profile is designed for instruments with red lasers. Researchers can evaluate far-red detector performance without relying exclusively on blue- or violet-laser excitation.

Dilution-Series Preparation

The 0.5% solids suspension can be diluted to prepare different working concentrations for concentration optimization, event-rate testing, threshold evaluation, and sample preparation studies.

The appropriate dilution should be determined experimentally for each flow cytometer and application.

Key Product Features

500nm Submicron Particle Size

The nominal 500nm diameter is suitable for research involving small-particle flow cytometry, fluorescence threshold development, and submicron detection workflows.

Far-Red Fluorescence

Excitation around 630nm and emission from 665 to 720nm provide compatibility with red-laser flow cytometry platforms and far-red fluorescence detectors.

Broad Emission Range

The broad emission profile allows researchers to evaluate different detector filters within the far-red region.

Stable Microsphere Suspension

The product is supplied as an aqueous suspension to support convenient dilution, mixing, and integration into laboratory workflows.

Suitable for Research and Product Development

RCY500NM-630 can support early-stage feasibility studies, assay optimization, instrument development, reagent development, and scale-up evaluation.

Bulk Manufacturing Capability

SHBC supports sample evaluation, pilot production, repeat orders, and bulk manufacturing for research organizations and reagent development companies.

Project-Based Customization

Particle concentration, packaging format, surface functionality, fluorescence intensity, particle size, and other project parameters may be discussed for qualified custom-development programs.

Flow Cytometry Applications

RCY500NM-630 Flow Red-Fluo Microspheres may be evaluated for the following research applications.

Small-Particle Flow Cytometry

Use the microspheres to develop acquisition and analysis methods for fluorescent submicron particles.

Potential studies include:

• Small-particle detection sensitivity

• Fluorescence-triggered event acquisition

• Scatter-versus-fluorescence comparison

• Electronic threshold optimization

• Background-event discrimination

• Sample dilution optimization

• Flow-rate comparison

• Gating strategy development

Extracellular Vesicle Method Research

The 500nm microspheres may be used as synthetic particles during the development of flow cytometry workflows for microvesicle- and extracellular-vesicle-related research.

They should not be treated as biological extracellular vesicle reference materials or certified size standards unless supported by the required lot-specific qualification.

Flow Cytometer Research and Development

Instrument manufacturers and research teams can use fluorescent microspheres when studying:

• Red-laser excitation performance

• Far-red detector sensitivity

• Optical alignment concepts

• Fluidic stability

• Event-rate performance

• Small-particle detection limits

• Detector-filter selection

• Data acquisition settings

The suitability of the product for formal instrument calibration must be independently validated.

Fluorescence Channel Evaluation

RCY500NM-630 can be used to evaluate detector channels that overlap the 665–720nm emission range.

Possible filter regions may include approximately:

• 660–680nm

• 665–695nm

• 670–710nm

• 680–720nm

• 690–735nm

Exact compatibility depends on the flow cytometer’s laser wavelength, detector sensitivity, dichroic mirrors, and bandpass filters.

Multiplex Flow Assay Development

Far-red fluorescent microspheres may be evaluated as one particle population in multiplex flow cytometry research.

Different particle sizes, fluorescence intensities, emission colors, or surface chemistries can potentially be combined to create distinguishable bead populations.

Multiplex performance must be optimized according to the instrument configuration, reporter fluorophores, compensation requirements, and assay design.

Fluorescent Particle Tracking

The microspheres may also support research involving:

• Particle transport

• Microfluidic flow studies

• Filtration research

• Particle uptake studies

• Fluorescence microscopy

• Fluidic-system evaluation

• Sample preparation research

Application suitability should be confirmed under the actual experimental conditions.

Recommended Laser and Detection Channels

RCY500NM-630 is designed for excitation at approximately 630nm.

It may be evaluated with red-laser flow cytometers using laser wavelengths including:

• 630nm

• 633nm

• 635nm

• 637nm

• 640nm

The expected emission is distributed between 665 and 720nm. A detector should therefore use an optical filter that overlaps a suitable portion of this range.

Potentially compatible far-red channels may include APC-like, far-red, red-laser fluorescence, or extended-red detector configurations. Channel names vary among instrument manufacturers, so filter specifications should be reviewed rather than relying only on the detector name.

Before starting a full study, confirm:

1. The instrument has a compatible red laser.

2. The emission filter overlaps 665–720nm.

3. The detector is sensitive enough for 500nm particles.

4. The instrument supports appropriate small-particle threshold settings.

5. The sheath fluid and sample buffer have low particulate background.

6. The selected concentration does not produce excessive coincident events.

RCY500NM-630 is not primarily designed for excitation with 405, 488, or 561nm lasers unless separate spectral testing confirms adequate excitation at those wavelengths.

Suggested Flow Cytometry Workflow

The following workflow is a general starting point and should be optimized for the intended instrument and research application.

1. Prepare the Instrument

Clean the fluidic system according to the instrument manufacturer’s recommendations.

Use clean, filtered sheath fluid and confirm that background events are acceptably low before analyzing the microspheres.

For submicron particle detection, cleanliness of the sheath fluid, sample tubes, buffers, and instrument fluidics is especially important.

2. Prepare a Buffer Blank

Run the same buffer used to dilute the microspheres.

Record the background event rate in the scatter and fluorescence channels before adding particles.

A buffer blank helps distinguish microsphere events from electronic noise, buffer particles, tube contamination, or carryover.

3. Resuspend the Microspheres

Allow the vial to reach the recommended working temperature before use.

Mix gently by inversion or controlled vortexing until the suspension appears uniform. A brief, mild bath-sonication step may be evaluated when necessary to redisperse settled particles.

Avoid harsh or prolonged sonication that could heat the sample or affect the formulation.

4. Prepare a Dilution Series

Prepare several working concentrations rather than using only one dilution.

A serial dilution can help determine the concentration that provides:

• A stable event rate

• Minimal coincident events

• Clear particle resolution

• Acceptable background separation

• Reproducible fluorescence intensity

Use clean, low-binding tubes and freshly filtered, particle-compatible buffer.

5. Select the Red Laser

Use a red laser close to the specified 630nm excitation wavelength.

A 633, 635, 637, or 640nm laser may also provide suitable excitation, but the fluorescence response should be experimentally confirmed.

6. Select the Far-Red Detector

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

Run the sample at low concentration first and adjust detector gain or voltage gradually to avoid signal saturation.

7. Optimize Threshold and Trigger Settings

For 500nm particles, conventional forward-scatter triggering may not provide the best sensitivity on every instrument.

Compare:

• Forward-scatter triggering

• Side-scatter triggering

• Fluorescence triggering

• Combined gating approaches

Record all acquisition settings so the method can be reproduced across experiments.

8. Control the Event Rate

Use a low or moderate sample flow rate during initial optimization.

A high particle concentration or excessive flow rate may increase coincident events, causing two or more particles to be detected as a single event.

Dilute the sample if the event rate is unstable or unexpectedly high.

9. Confirm Reproducibility

Repeat the measurement using independently prepared dilutions.

Evaluate:

• Event count stability

• Fluorescence median or mean

• Distribution width

• Background separation

• Scatter position

• Run-to-run consistency

Quality Control and Batch Consistency

For flow cytometry reagent development and bulk manufacturing, consistency between production batches is an important consideration.

Depending on project requirements, quality evaluation may include:

• Particle-size characterization

• Particle-size distribution

• Suspension appearance

• Solids-content verification

• Fluorescence excitation testing

• Fluorescence emission testing

• Relative fluorescence-intensity comparison

• Flow cytometry histogram evaluation

• Dispersion and aggregation assessment

• Accelerated or real-time stability evaluation

• Batch-to-batch comparison

Available release specifications and testing methods should be agreed upon before bulk production.

For projects requiring quantitative fluorescence standards, certified particle-size standards, MESF assignments, traceability, or instrument-calibration claims, additional qualification and documentation are required. Standard RCY500NM-630 microspheres should not be described as certified calibration beads unless the relevant certification has been completed.

Bulk Supply and Custom Manufacturing

SHBC provides fluorescent microsphere manufacturing support for biotechnology companies, flow cytometry laboratories, reagent developers, instrument manufacturers, and research organizations.

Our typical cooperation process can include:

Research Sample Evaluation

Small quantities can be evaluated for fluorescence intensity, detector compatibility, particle dispersion, background separation, and assay feasibility.

Pilot-Scale Development

Pilot batches can support method optimization, formulation screening, stability testing, and initial product-development studies.

Bulk Production

After technical specifications are confirmed, repeat production and bulk quantities can be arranged according to the project forecast.

Custom Project Discussion

Available customization may include:

• Alternative particle diameters

• Different fluorescence colors

• Adjusted fluorescence intensity

• Alternative solids concentrations

• Project-specific packaging

• Surface-functionalized microspheres

• Carboxyl, amino, or other surface options

• Biomolecule-coupling development

• Lot-specific quality-control requirements

The feasibility of each customization depends on the requested particle size, fluorescence spectrum, surface chemistry, concentration, packaging, and order quantity.

Handling and Storage Recommendations

For best performance:

• Store the microspheres protected from light.

• Refrigerated storage at 2–8°C is generally recommended unless otherwise stated in the product documentation.

• Do not freeze the suspension unless freeze-thaw stability has been specifically validated.

• Keep the vial tightly closed when not in use.

• Avoid prolonged exposure to strong laboratory light.

• Mix thoroughly before sampling.

• Use clean pipette tips and low-particle-background tubes.

• Prevent microbial or particulate contamination.

• Avoid repeated transfer between containers.

• Do not allow the suspension to dry.

• Record the lot number and opening date for traceability.

Before use, inspect the suspension for unexpected aggregation, color change, contamination, or irreversible settling.

Storage conditions, preservative information, shelf life, and transport requirements should be confirmed using the product label, technical data sheet, safety data sheet, or certificate of analysis supplied with the order.

Frequently Asked Questions

What is RCY500NM-630?

RCY500NM-630 is a 500nm far-red fluorescent microsphere suspension with 0.5% solids. It is designed for excitation around 630nm and emits fluorescence between 665 and 720nm.

Which flow cytometry laser should be used?

A red laser near 630nm is recommended. Instruments with 633, 635, 637, or 640nm lasers may also be suitable, but excitation efficiency should be confirmed experimentally.

Which detector should be selected?

Select a detector filter that overlaps the 665–720nm emission range. Detector names vary among flow cytometer manufacturers, so the actual bandpass-filter specification should be checked.

Can these microspheres be detected by forward scatter?

Detection depends on the flow cytometer’s optical design and small-particle sensitivity. Some conventional cytometers may have limited scatter sensitivity at 500nm. Fluorescence-triggered detection may provide better event identification during submicron particle studies.

Can RCY500NM-630 be used for extracellular vesicle research?

The microspheres may be evaluated as synthetic particles during extracellular vesicle or microvesicle flow cytometry method development. They are not biological extracellular vesicles and should not automatically be treated as certified size or concentration standards.

Are these certified calibration beads?

Not unless the supplied product documentation specifically states that certification or traceability is included. Standard fluorescent microspheres can support method development and comparative studies, but formal calibration claims require additional qualification.

Why is a dilution series recommended?

A dilution series helps identify an appropriate particle concentration, reduce coincident events, optimize the event rate, and distinguish genuine particles from background events.

Can the microspheres be used in multiplex flow cytometry?

They may be evaluated as a far-red particle population in multiplex research. Compatibility with other bead populations and reporter fluorophores must be tested using the intended instrument and compensation strategy.

Can the particle surface be functionalized?

Project-specific surface chemistry may be discussed. Depending on the application, carboxyl, amino, or other functionalized fluorescent microspheres may be available through custom development.

Can antibodies or proteins be coupled to the microspheres?

Biomolecule coupling requires a compatible surface functional group and an optimized coupling process. Please specify the desired biomolecule, surface chemistry, assay format, particle size, and target application when requesting a customized product.

Are other particle sizes available?

Different nanometer- and micrometer-scale fluorescent microsphere sizes can be discussed for flow cytometry, fluorescence microscopy, immunoassay, particle tracking, and instrument-development projects.

Is bulk production available?

Yes. RCY500NM-630 is intended for research evaluation, pilot-scale development, and bulk supply. Packaging format, annual demand, QC requirements, and delivery schedule can be discussed during project evaluation.

What information should be provided when requesting a quotation?

Please provide:

• Product catalog number

• Required quantity

• Expected annual demand

• Preferred package size

• Flow cytometer laser wavelength

• Detector-filter configuration

• Required surface chemistry

• Target fluorescence intensity

• Application description

• Required quality documentation

• Delivery destination

Request Samples or a Bulk Quotation

RCY500NM-630 500nm Flow Red-Fluo Microspheres provide a far-red fluorescent particle option for red-laser flow cytometry research, submicron detection method development, fluorescence channel evaluation, instrument development, and flow-based assay research.

For sample evaluation or bulk production, please provide your instrument configuration, application, required quantity, preferred packaging, and quality-control requirements.

Product: 500nm Flow Red-Fluo Microspheres
Catalog Number: RCY500NM-630
Particle Diameter: 500nm
Excitation: 630nm
Emission: 665–720nm
Solids Content: 0.5%
Application: Flow Cytometry Research
Supply: Samples, Pilot Batches, and Bulk Production
Use: Research Use Only. Not for diagnostic or therapeutic use.