Cellular Stains

Membrane Stains

Membrane & Cell Surface Stains Comparison ... p. 4

CellBrite™ Steady Membrane Dyes for Live Cell Imaging ... p. 4

Fixable Surface Stains: CellBrite™ Fix & MemBrite™ Fix ... p. 5

CellBrite™ Lipophilic Membrane Dyes for Live or Fixed Cells ... p. 5

Lectin Conjugates ... p. 6

LipidSpot™ Lipid Droplet Stains ... p. 6

Labeled Phospholipids ... p. 6

Cytoplasmic Stains and Cell Division Tracking

ViaFluor® SE Cell Proliferation Dyes ... p. 3

Calcein AM Cell Viability Assay ... p. 3

Organelle Stains

Nuclear Stains ... p. 7

MitoView™ Mitochondrial Stains ... p. 8

LysoView™ Lysosomal Stains ... p. 9

Cytoskeleton Probes ... p. 10

Endosomal Tracers ... p. 11

Overview of Organelle & Cytoskeleton Stains ... p. 2

2 • www.biotium.com

Overview of Organelle &

Cytoskeleton Stains

Cytoskeleton

• Live cell microtubule stains (p. 10)

• Phalloidin conjugates for xed cells (p. 10)

Mitochondria

Membrane-permeant dyes that

accumulate in mitochondria due

to membrane potential &

lipophilicity (p. 8)

Vesicle Tracking

• Fluorescent toxins and ligands for receptor-mediated endocytosis (p. 11)

• Fluorescent dextrans for uid phase tracing (p. 11)

Nucleus

Blue, green, and red stains with

various properties for labeling

xed, dead, or live cells (p. 7)

Cytoplasm

• Stable,covalentlivecellcytoplasmic

stainsforcelldivisionanalysisbyow

ortrackingcellsinculture(p.3)

• Non-covalentcytoplasmstainsforcell

viability&dyeeuxassays(p.3)

Lysosomes

Membrane-permeant, pH-sensitive

dyes that accumulate and uoresce

in the acidic environment of

lysosomes (p. 9)

Membrane & Cell Surface

• Cell surface stains for long-term imaging of live cells (p. 4)

• Lipophilic carbocyanine dyes for live or xed cells (p. 5)

• Covalent membrane & surface stains for xable

live-cell staining (p. 5)

• Lectin conjugates for staining cell surface

glycoproteins in live or xed cells (p. 6)

Lipid Droplets

Neutral lipid stains for

live or xed cells (p. 6)

www.biotium.com • 3

ViaFluor® SE Cell Proliferation Kits use amine-reactive dyes to covalently label cells throughout the cell cytoplasm and intracellular compartments. While developed

for cell division tracking by ow cytometry, stable whole-cell labeling is also useful for imaging cell morphology, or to track cell populations in mixed co-culture

experiments (Fig. 1). In addition, the labeling is covalent and can therefore withstand xation and permeabilization for downstream immunostaining.

ViaFluor® CFSE is the classic cell proliferation dye, detected in the FITC channel. Biotium created ViaFluor® 488 as an improved alternative to CFSE that is less

toxic, less leaky, and has a greater tolerance for xation. We also offer blue uorescent ViaFluor® 405 for the Pacic Blue® channel which provides bright and

stable staining with low toxicity.

ViaFluor® SE Cell Proliferation Kits

Cytoplasmic Stains and Cell Division Tracking

Cat. # Product Ex/Em (nm)

30068 ViaFluor® 405 SE Cell Proliferation Kit 408/452

30086 ViaFluor® 488 SE Cell Proliferation Kit 493/532

30050 ViaFluor® CFSE Cell Proliferation Kit 495/519

ViaFluor® SE Cell Proliferation Dyes

Figure 1. Adherent HeLa cells and MCF-7 cells were labeled

with ViaFluor® 488 (green) and ViaFluor® 405 (blue),

respectively. The cells were then trypsinized, mixed, plated,

and grown in co-culture for four days.

ViaFluor® SE Cell Proliferation Kit Features

• Non-toxic dyes for covalent cytoplasm labeling

• Suitable for long-term imaging

• Toleratesxationandpermeabilization

• ViaFluor® 405 SE and ViaFluor® 488 SE offer

better performance and less toxicity than CFSE

Calcein AM is a widely used green uorescent live cell stain. The compound is initially a non-uorescent, membrane permeable compound. Once inside live cells,

cytoplasmic esterase activity converts calcein AM to the green uorescent, membrane-impermeant compound calcein that lls the entire cell (Fig. 2). We offer

high-purity Calcein AM as a solid or as a ready-to-use solution in the Calcein AM Cell Viability Assay Kit. Because calcein is only retained in cells with intact plasma

membranes, the Calcein AM Cell Viability Assay is a true live-cell endpoint assay that is compatible with microplate readers. Staining is fast and simple, requiring

only 30 minutes.

The Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells pairs calcein AM with the red dead cell stain Ethidium Homodimer III, to detect live

and dead cells by microplate reader, ow cytometry, or microscopy (Fig. 3).

Calcein AM for Cell Viability

Figure 2. Live HeLa cells labeled with Calcein AM.

Figure 3. HeLa cells stained with the Viability/Cytotoxicity

Assay Kit for Animal Live & Dead Cells. Live cells are stained

green, dead cells are stained red.

Cat. # Product Ex/Em (nm)

80011 Calcein AM

494/517 (hydrolyzed product)

30026 Calcein AM Cell Viability Assay Kit

30002

Viability/Cytotoxicity Assay Kit for Animal

Live & Dead Cells

494/517 (Calcein AM)

522/593 (EthD-III)

Cytosolic Tracers

Pacic Blue is a registered trademark of Thermo Fisher Scientic.

4 • www.biotium.com

CellBrite™ Steady Features

• Rapid & uniform staining of live cell surface

membranes for 24 hours or longer

• Optional Enhancer masks intracellular signal

for selective imaging of cell surface

• Dye colors from blue to near-IR, with

STORM-compatible options

Membrane & Cell Surface Stains

CellBrite™ Steady Membrane Staining Kits

CellBrite™ Steady Membrane Staining Kits are unique uorescent membrane

probes that allow uorescence imaging of cell surface for up to several days

in culture. Unlike non-covalent lipophilic membrane stains that are rapidly lost

from the cell surface by endocytosis after labeling, CellBrite™ Steady Dyes

equilibrate between intracellular compartments and the plasma membrane.

Therefore, cells retain uniform cell surface and intracellular staining for 24

hours or longer (Fig. 1). CellBrite™ Steady Enhancer is an optional reagent

included in the kits that can be used to mask intracellular uorescence of

CellBrite™ Steady Dyes, for more selective visualization of cell boundaries.

Washing optional for confocal imaging, required for epiuorescence.

Staining is retained immediately after formaldehyde xation, but dye redistributes over time; if cells must be xed, image within 24 hours

Product Line Colors Protocol

Stain live

cells

Stain xed

cells

Fixable

Staining

Applications/Notes

CellBrite™ &

CellBrite™ NIR

Cytoplasmic

Membrane Dyes

Blue to

Near-IR

• Stain in complete medium ~20 min

• Washing required

• Staining lost from cell surface over

time by internalization

Yes Yes (PFA)

Yes (PFA)

Does not

tolerate

detergent or

MeOH xation

• Lipophilic carbocyanine dyes

• Non-toxic with minimal dye transfer between cells

• Suitable for live-cell surface staining, transplantation, or co-culture

• Can stain cell before or after PFA-xation

CellBrite™ Fix

Membrane Stains

Green, Red,

Far-Red

• Stain in buffer 15 min

• Washing optional

Yes No

Yes

Tolerates MeOH

xation &

detergent

• Fixable membrane stains that tolerate permeabilization

• For labeling cell surface before xation & immunouorescence

• Non-toxic, but covalent modication of cell surface may affect function

MemBrite™ Fix

Cell Surface

Stains

Blue to

Near-IR

• Treat with prestain solution 5 min

• Stain in buffer 5 min

• Washing required

Yes No

Yes

Tolerates MeOH

xation &

detergent

• Fixable membrane stains that tolerate permeabilization

• For labeling cell surface before xation & immunouorescence

• STORM-compatible dye options

• Non-toxic, but covalent modication of cell surface may affect function

CF® Dye WGA

& Other Lectin

Conjugates (p. 6)

Blue to

Near-IR

• Stain in complete medium 10 min

• Washing recommended

Yes Yes

Yes

Tolerates MeOH

xation &

detergent

• Stain cells or tissue sections (staining is cell-type and tissue-dependent)

• Many dye options, including STORM-compatible dyes

• Binds to specic glycoproteins; staining may be toxic

CellBrite™ Steady

Membrane Dyes

Blue to

Near-IR

• Stain in complete medium 30 min to

several days

• No wash staining for confocal imaging

• Equilibrates between cell surface and

intracellular compartments

Yes No

Yes (PFA)

Does not

tolerate

detergent or

MeOH

• Unique probes for stable multi-day imaging of the cell surface membrane

• Non-toxic, uniform, & even cell staining

• Optional enhancer masks intracellular signal

• STORM-compatible dye options

Membrane & Cell Surface Stains Comparison

Biotium offers a wide selection of highly uorescent and photostable stains to visualize cell boundaries and morphology in multi-color staining experiments. Our

novel CellBrite™ Fix and MemBrite™ Fix Membrane Stains are unique cell surface stains that can tolerate detergent permeabilization and methanol xation.

CellBrite™ Steady dyes are excellent for long-term imaging of cell surface membranes in live cells. Our original CellBrite™ Cytoplasmic Membrane Stains are

lipophilic dyes for simple, non-toxic, and stable labeling of membranes in live or xed cells. We also offer a several lectins conjugated to our bright and photostable

CF® dyes for cell surface staining after xation or permeabilization (see p. 6).

DiD Staining

30 minutes 3 hours

CellBrite™ Steady 650 + Enhancer

30 minutes 24 hours

Figure 1. Comparison of lipophilic carbocyanine dye DiD and CellBrite™ Steady

in live HeLa cells. DiD staining is heterogeneous and rapidly internalized into

intracellular compartments. In contrast, CellBrite™ Steady 650 gives even staining

of cell surface that can be clearly imaged after 24 hours in culture with the use

of Enhancer.

www.biotium.com • 5

CellBrite™ Fix and MemBrite™ Fix Features

• Toleratepermeabilizationandxation

• Stable and non-cytotoxic

• Simple and rapid staining protocols

• MemBrite™ Fix-ST dyes for STORM

• Suitable for mammalian cells, yeast, & bacteria

While other lipophilic membrane dyes such as DiB, DiO and DiI can be xed

with formaldehyde, they are not compatible with detergent permeabilization

or methanol xation. To address these issues Biotium developed the novel

CellBrite™ Fix and MemBrite™ Fix dyes. These are unique dyes that

accumulate at the cell surface and bind covalently to membrane proteins for

stable labeling. Both types of dyes can tolerate permeabilization and methanol

xation (Fig. 1), allowing membrane staining to be combined with intracellular

labeling with antibodies. Moreover, CellBrite™ Fix and MemBrite™ Fix dyes

offer greater water solubility compared to lipophilic carbocyanine dyes like

DiO and DiI, resulting in much more uniform staining. The dyes are also non-

cytotoxic and provide stable labeling that does not transfer readily between

cells, allowing co-culture experiments.

CellBrite™ Fix and MemBrite™ Fix

Membrane Dyes That Tolerate Permeabilization

Figure 1. CellBrite™ Fix and MemBrite™ Fix label the cell surface more evenly than

other lipophilic membrane stains, and tolerate xation and permeabilization. Because

CellBrite™ Fix and MemBrite™ Fix labeling is covalent, it does not redistribute after

permeabilization, unlike lipophilic carbocyanine dye DiI. Live HeLa cells were labeled with DiI,

CellBrite™ Fix 555, or MemBrite™ Fix 568/580. Cells were xed with 4% paraformaldehyde

(PFA) (left column), followed by permeabilization with 0.1% Triton® X-100 (center column), or

xed with methanol (MeOH) (right column).

DiI Staining

CellBrite™ Fix 555

MemBrite™ Fix 568/580

MeOH x

PFA x +

Triton® permPFA x

CellBrite™ Cytoplasmic Membrane Dyes

CellBrite™ Cytoplasmic Membrane dyes are lipophilic carbocyanine dyes like

DiI and DiO, which have been used extensively for cell tracing and tracking.

Unlike other membrane stains, these dyes can be used for selective plasma

membrane staining after cells are xed and permeabilized. We offer a selection

of dyes with uorescence ranging from blue to near-infrared. We also offer

CellBrite™ NIR dyes which are suitable for in vivo near-IR imaging in small

animals. Please visit our website to view our full selection of stand-alone

lipophilic carbocyanine dyes including DiO, DiI, and DiR.

Cat. # Product Ex/Em (nm)

30092 MemBrite™ Fix 405/430 Cell Surface Staining Kit 405/430

30093 MemBrite™ Fix 488/515 Cell Surface Staining Kit 488/515

30094 MemBrite™ Fix 543/560 Cell Surface Staining Kit 543/560

30095 MemBrite™ Fix 568/580 Cell Surface Staining Kit 568/580

30096 MemBrite™ Fix 594/615 Cell Surface Staining Kit 594/615

30097 MemBrite™ Fix 640/660 Cell Surface Staining Kit 640/660

30098 MemBrite™ Fix 660/680 Cell Surface Staining Kit 660/680

30099 MemBrite™ Fix 680/700 Cell Surface Staining Kit 680/700

30101 MemBrite™ Fix-ST 650/665 Cell Surface Staining Kit 650/665

30102 MemBrite™ Fix-ST 667/685 Cell Surface Staining Kit 667/685

30103 MemBrite™ Fix-ST 681/698 Cell Surface Staining Kit 681/698

30104 MemBrite™ Fix-ST 755/777 Cell Surface Staining Kit 755/777

MemBrite™ Fix Cell Surface Labeling Kits

Cat. # Product Ex/Em (nm)

30090 CellBrite™ Fix 488 Membrane Stain 480/513

30088 CellBrite™ Fix 555 Membrane Stain 542/571

30089 CellBrite™ Fix 640 Membrane Stain 638/667

CellBrite™ Fix Membrane Labeling Kits

Cat. # Product Ex/Em (nm)

30024 CellBrite™ Blue Cytoplasmic Membrane Staining Kit 366/441

30021 CellBrite™ Green Cytoplasmic Membrane Labeling Dye 484/501

30022 CellBrite™ Orange Cytoplasmic Membrane Labeling Dye 549/565

30023 CellBrite™ Red Cytoplasmic Membrane Labeling Dye 644/665

30070 CellBrite™ NIR680 Membrane Labeling Dye 683/724

30077 CellBrite™ NIR750 Membrane Labeling Dye 748/780

30078 CellBrite™ NIR770 Membrane Labeling Dye 767/806

30079 CellBrite™ NIR790 Membrane Labeling Dye 786/820

CellBrite™ & CellBrite™ NIR Cytoplasmic Membrane Dyes

Cat. # Product Ex/Em (nm)

30105 CellBrite™ Steady 405 Membrane Staining Kit 406/428

30106 CellBrite™ Steady 488 Membrane Staining Kit 505/529

30107 CellBrite™ Steady 550 Membrane Staining Kit 562/579

30108 CellBrite™ Steady 650 Membrane Staining Kit 656/676

30109 CellBrite™ Steady 685 Membrane Staining Kit 686/708

CellBrite™ Steady Membrane Labeling Kits

Membrane & Cell Surface Stains

6 • www.biotium.com

Lectin Conjugates and Lipid Stains

Intracellular lipid droplets are cytoplasmic organelles involved in the storage

and regulation of triglycerides and cholesterol esters. LipidSpot™ dyes are

uorogenic neutral lipid stains that rapidly stain lipid droplets with minimal

background staining of cellular membranes or other organelles (Fig. 2). The

protocol is simple and convenient, requiring as little as 30 minutes for incubation

before or after xation with no wash step required. LipidSpot™ dyes are

available in green or far-red uorescence with compatible detection channels

for FITC, Texas Red®, or Cy®5. LipidSpot™ 488 has green fluorescence for

the FITC channel and has been validated in super-resolution imaging by SIM.

LipidSpot™ 610 has red/far-red fluorescence than can be detected in the Texas

Red® or Cy® 5 channel.

LipidSpot™ Lipid Droplet Stains

Texas Red is a registered trademark of Thermo Fisher Scientic. Cy dye is a registered trademark

of GE Healthcare.

LipidSpot™ Lipid Droplet Stains

Cat. # Product Ex/Em (nm)

70065 LipidSpot™ 488 Lipid Droplet Stain, 1000X in DMSO 420/511

70069 LipidSpot™ 610 Lipid Droplet Stain, 1000X in DMSO 610/663

Cat. # Product Ex/Em (nm)

60022 Biotin-DHPE N/A

60023 Biotin-X-DHPE N/A

60024 Fluorescein-DHPE 496/519

60025 NBD-PE 463/536

60028 TRITC-DHPE 540/566

60026 Rhodamine-DHPE 560/581

60027 Texas Red®-DHPE 582/601

Phospholipid Probes

These membrane probes are derived from natural phospholipids by

modifying the head group with a uorescent dye or biotin. The probes are

useful for studying vesicle trafcking and membrane fusion. Red uorescent

phospholipids like TRITC-, Rhodamine-, and Texas Red®-DHPE have been

used as uorescence acceptors in combination with NBD-DHPE in membrane

fusion assays.

Labeled Phospholipids

LipidSpot™ Features

• Rapid staining of lipid droplets

• Compatiblewithliveorxedcells

• Simple protocol with no wash step

• Minimal background staining

• Detect in FITC, Texas Red®, or Cy®5 channels

CF ® Dye Labeled Lectins

Dye Ex/Em (nm) Con A PNA WGA

CF®350 347/448 29015 --- 29021

CF®405S 404/431 29075 --- 29027

CF®405M 408/452 29074 --- 29028

CF®488A 490/515 29016 29060 29022

CF®532 527/558 --- --- 29064

CF®555 555/565 --- --- 29076

CF®568 562/583 --- 29061 29077

CF®594 593/614 29017 29062 29023

CF®633 630/650 29018 --- 29024

CF®640R 642/662 29019 29063 29026

CF®680 681/698 29020 --- 29029

CF®680R 680/701 --- --- 29025

CF®750 755/777 29080 --- ---

CF®770 770/797 29058 --- 29059

CF® Dye Lectins

Lectins are carbohydrate binding proteins that recognize specic sugar moieties

on cell surface glycoproteins. They can be used to stain the cell surface of live

cells; and withstand xation and permeabilization. When cells are xed and

permeabilized before staining, uorescent lectins stain both cell surface and

organelles in the secretory pathway. Biotium offers CF® dye lectin conjugates

of wheat germ agglutinin (WGA) (Fig. 1), concanavalin A (Con A), and peanut

lectin from Arachis hypogaea (PNA) in a variety of colors. Con A conjugates are

used to stain the yeast cell wall and WGA conjugates are used to stain yeast

bud scars. WGA may also be used as a live-cell Gram stain for bacteria.

Figure 1. Live HeLa cells stained with CF®488A

WGA (green) and Hoechst 3342 (blue).

Figure 2. Oleic acid-treated HeLa cells were

xed with formaldehyde and stained with

LipidSpot™ 488 (green), CF®594 WGA (red),

and Hoechst (blue).

www.biotium.com • 7

Nuclear Stains

NucSpot® Nuclear Stains for Live or Fixed Cells

NucSpot® Live Cell Nuclear Stains are cell-permeable DNA dyes that stain

nuclei in live or fixed cells. Staining is specific, non-toxic, and does not

require a wash step, so staining can be continuously monitored for several

days. NucSpot® Live 488 has green uorescence and NucSpot® Live 650 has

far-red uorescence for the Cy®5 channel (Fig. 1). NucSpot® Live 650 is also

suitable for super-resolution imaging techniques such as STORM or SIM.

Figure 2. Formaldehyde xed and detergent permeabilized HeLa

cells stained with RedDot™2 (A) or Draq7™ (B). Actin is stained with

CF®488A phalloidin (green).

A. RedDot™2 B. Draq7™

A. NucSpot® Live 488 B. NucSpot® Live 650

Figure 1. Live HeLa cells stained with 1X NucSpot® Live 488 (A)

or 1X NucSpot® Live 650 (B).

Draq7 is a trademark of Biostatus, Ltd. Cy Dye is a registered trademark of GE Healthcare.

In-Cell Western is a trademark of LI-COR® Biosciences. SYTOX, TOTO, and TO-PRO are

registered trademarks of Thermo Fisher Scientic.

*With DNA

Cat. # Product Ex/Em (nm)

40081 NucSpot® Live 488 Nuclear Stain, 1000X in DMSO 500/515

40082 NucSpot® Live 650 Nuclear Stain, 1000X in DMSO 650/675

40083 NucSpot® 470 Nuclear Stain, 1000X in DMSO 460/546

40060 RedDot™1 Far-Red Nuclear Stain, 200X in Water 662/694*

40061 RedDot™2 Far-Red Nuclear Stain, 200X in DMSO 665/695*

40011 DAPI 358/461*

40044 Hoechst 33258, 10 mg/mL in H

O 352/461

40046 Hoechst 33342, 10 mg/mL in H

O 350/461

Nuclear Stains

RedDot™1 and RedDot™2 are far-red nuclear counterstains for the Cy®5

channel. RedDot™1 rapidly and specically stains nuclei in live cells, and can

be used for cell cycle analysis by ow cytometry or for cell normalization by

In-Cell Western™. It has also been used to stain nuclei in live atworms.

RedDot™2 is membrane impermeant and can be used to selectively stain dead

cells, or as a nuclear counterstain for xed cells. RedDot™2 is functionally

similar to Draq7™, but with better nuclear specicity in xed cells and does not

require a blocking step for nuclear-specic staining (Fig. 2).

RedDot™1 and RedDot™ 2 Far-Red Nuclear Stains

NucSpot® 470 is a cell membrane-impermeant green fluorescent DNA

stain. While other green nucleic acid stains like TOTO®, TO-PRO®, or

SYTOX® dyes stain both the nucleus and cytoplasm, NucSpot® 470

specifically stains the nucleus of fixed and permeabilized cells. It can also

be used to selectively stain dead cells in living cultures. It can be imaged

using standard settings for FITC, and is an excellent match for instruments

with blue LED excitation.

NucSpot® 470 Nuclear Stain for Dead or Fixed Cells

NucSpot® Features

• Excellentnuclearspecicity,noRNaseor

washing required

• Greenorfar-reduorescence

• NucSpot® Live Stains have low toxicity and are

suitable for long-term imaging

• NucSpot®470stainsdeadorxedcells

RedDot™ 1 Features

• Cell-permeable, for staining live cells

• Canbeusedforcellcycleanalysisbyow

cytometry

RedDot™ 2 Features

• Forxedcellsortissues,orselectivedeadcell

staining

• Morenuclear-specicthanDraq5™or

Draq7™, with no RNase or wash required

We offer a number of widely used nuclear specic dyes for microscopy

analysis. This includes DAPI and Hoechst, commonly used for live or xed cell

nuclear staining. We also offer antibody conjugates that label nuclei, nuclear

membranes, and nucleoli.

Visit our website at www.biotium.com to view our full selection of dead cell

nucleic acid stains like propidium iodide (PI), 7-AAD, Ethidium Homodimer

III, dimeric cyanine dyes (also known as TOTO® and TO-PRO®), as well as

mounting media with DAPI.

Other Nucleic Acid Dyes

8 • www.biotium.com

Mitochondrial Stains

500

1000

1500

2000

Control

CCCP

Staurosporine

Geometric mean (FL4-H)

Figure 2. Flow cytometry analysis of Jurkat cells treated

with CCCP to depolarize the mitochondrial membrane or

staurosporine to induce apoptosis, resulting in decreased

MitoView™ 633 staining.

Figure 1. Live HeLa cells stained with MitoView™ 405 (A), MitoView™ Green (B),

MitoView™ 633 (C), or MitoView™ 720 (D).

A. MitoView™ 405 B. MitoView™ Green

C. MitoView™ 633 D. MitoView™ 720

MitoView™ 633 also has red uorescence in the Cy®3/rhodamine channel. It is not recommended

for imaging with other red uorescent probes.

While optimal for Cy®7 settings, MitoView™ 720 is bright enough to be imaged in the Cy®5

channel, and can be combined with visible red uorescent probes.

Dyes with partial mitochondrial membrane potential dependence localize to the cytoplasm after

mitochondrial depolarization, but still retain uorescence.

Cat. # Product Ex/Em (nm)

Potential-

dependent?

70070 MitoView™ 405 398/440 Partial

70054 MitoView™ Green 490/523 No

70055 MitoView™ 633 622/648

Yes

70075 MitoView™ 650 644/670 Partial

70068 MitoView™ 720 720/758

Partial

MitoView™ Dyes

Cat. # Product Features

30062

NucView® 488 and MitoView™

633 Apoptosis Kit

Two color detection of caspase-3 activity

and mitochondrial potential

30001

JC-1 Mitochondrial Membrane

Potential Detection Kit

Two-color detection of mitochondria

polarization/depolarization

Assay Kits

Cy Dye is a registered trademark of GE Healthcare.

MitoView™ Features

• Rapid, no-wash, live cell mitochondria stains

• Bright, photostable, and non-toxic

• Availableinvecolorswithblue,green,far-red,

or near-IR emission

• Options for mitochondrial potential-dependent

or -independent staining

MitoView™ Dyes

MitoView™ dyes are uorogenic mitochondrial stains for live cells (Fig. 1).

The dyes rapidly stain mitochondria without a wash step, and are non-toxic

for live-cell imaging. They are available with blue, green, far-red, and near-

infrared uorescence. MitoView™ 633 can be used to monitor mitochondrial

membrane potential by microscopy or ow cytometry (Fig. 2). We also offer

MitoView™ Green, a potential-independent mitochondrial dye that can be

imaged following mitochondrial depolarization, or after xation. MitoView™ dyes

stain mitochondria in yeast, and also stain bacteria (gram-positive and gram-

negative).

More Mitochondrial Dyes

Cat. # Product Ex/Em (nm)

Potential-

dependent?

70014 JC-1, Iodide Salt

510/527; 585/590* Ratiometric*

70011 JC-1, Chloride Salt

70010 Rhodamine 123 505/534 Yes

70016 TMRE

549/574 Yes

70005 TMRE, 2 mM in DMSO

70017 TMRM 548/573 Yes

70018 DASPEI 461/589 Yes

70015 DiIC

(5) 638/658 Yes

70012 Nonyl Acridine Orange (NAO) 495/522 No

*JC-1 forms red uorescent aggregates in polarized mitochondria, and green uorescent monomers

in cytoplasm

Other Mitochondrial Dyes and Antibodies

We offer a wide selection of classic mitochondrial membrane-potential dyes

including Rhodamine 123 and TMRE/TMRM. JC-1 dye can be used for

ratiometric measurements of mitochondrial potential, while NAO is a green

potential-independent dye that binds cardiolipin phosphoprotein in mitochondria.

For xed cells and tissues we recommend using mitochondrial localized

antibodies. We offer several mitochondrial localized primary antibodies

conjugated to our bright and photostable CF® dyes. Visit our website at

www.biotium.com to learn more.

www.biotium.com • 9

Lysosomal Stains

LysoTracker is a registered trademark of Thermo Fisher Scientic

Cy Dye is a registered trademark of GE Healthcare.

Figure 2. pH dependence of LysoView™ 633 uorescence emission.

625 650 675 700 725

Emission

Wavelength (nm)

pH 2

pH 3

pH 4

pH 5

pH 6

pH 7

pH 8

pH 9

pH ≤ 5

pH 6

pH 7

pH 8

pH 9

Figure 1. Live HeLa cells stained with LysoView™ 405 (A), LysoView™ 488 (B),

LysoView™ 540 (red) and Hoechst 33342 (blue) (C), or LysoView™ 650 (magenta)

and MitoView™ Green (D).

C. LysoView™ 540 D. LysoView™ 650

A. LysoView™ 405 B. LysoView™ 488

Cat. # Product Ex/Em (nm)

70061 LysoView™ 405, 1000X in DMSO 318, 400/464

70061 LysoView™ 488, 1000X in DMSO 506/532

70061 LysoView™ 540, 1000X in DMSO 541/634*

70058 LysoView™ 633 (1000X)** 634/659*

70059 LysoView™ 650, 1000X in DMSO 650/675

*pH ≤ 5

**After reconstitution

LysoView™ Dyes

LysoView™ dyes are uorescent stains for imaging lysosome localization and morphology in live cells. LysoView™ dyes belong to a family of lysosomotropic

dyes with weakly basic amine groups that accumulate in acidic organelles. LysoView™ dyes are available with blue, green, red, and far-red uorescence (Fig. 1).

Red uorescent LysoView™ 540 and far-red uorescent LysoView™ 633 dyes exhibit pH-dependent uorescence (Fig. 2), resulting in specic lysosomal staining

without a wash step. LysoView™ 488 has been validated for super-resolution imaging by SIM, while LysoView™ 650 is compatible with SIM and STED.

LysoView™ Features

• Fluorescent imaging of lysosomes in live cells

• Highlyspecic,no-washstainingofacidic

organelles

• Dyes available for DAPI, FITC, Cy®3, and

Cy®5 channels

• LysoView™ 650 compatible with SIM and

STED imaging

• Options for yeast and bacteria staining

Visit website for details

Figure 3. LysoView™ 633 compared to LysoTracker® Deep Red. Live HeLa

cells were stained for 10 minutes at 37

C with 1X LysoView™ 633 or 50 nM

LysoTracker® Deep Red (Thermo Fisher Scientic). LysoView™ 633 (A) showed

more specic lysosomal staining with less cytoplasmic staining compared to

LysoTracker® Deep Red (B).

A. LysoView™ 633 B. LysoTracker® Deep Red

10 • www.biotium.com

A. ViaFluor® 405, B. ViaFluor® 488

C. ViaFluor® 647 D. ViaFluor® 647

Figure 1. Live HeLa cells stained with ViaFluor® 405 microtubule stain (blue) and

LysoView™ 540 (red) (A). HeLa cells stained with ViaFluor® 488 microtubule stain

(B). Live MCF-7 cells stained with ViaFluor® 647 microtubule stain (C). Dividing HeLa

cell stained with ViaFluor® 647 microtubule stain showing mitotic spindle (magenta).

DNA is stained with Hoechst 33342 (blue) (D).

Figure 2. HeLa cells stained with CF®405M

phalloidin (blue) and RedDot™2 Far-Red Nuclear

Stain (magenta).

Cat. # Product Ex/Em (nm)

70064-T

ViaFluor® 405 Live Cell Microtubule Stain,

1000X in DMSO

408/452

70064

70062-T

ViaFluor® 488 Live Cell Microtubule Stain,

1000X in DMSO

500/515

70062

70063-T

ViaFluor® 647 Live Cell Microtubule Stain,

1000X in DMSO

650/675

70063

ViaFluor® Live Cell Microtubule Stains

Alexa Fluor, Texas Red, and DyLight are registered trademarks of Thermo Fisher Scientic. Cy

Dye is a registered trademark of GE Healthcare. TAXOL is a registered trademark of Bristol-Myers

Squibb.

ViaFluor® Live Cell Microtubule Stains are simple, rapid, and sensitive

stains for imaging the microtubule cytoskeleton in live cells (Fig.1). The

stains are uorescent, cell-permeable TAXOL® (paclitaxel) probes which

label the cytoskeleton by binding to polymerized tubulin. Though TAXOL® is

known for stabilizing microtubules and inhibiting mitosis, ViaFluor® Live Cell

Microtubule Stains are less disruptive of microtubule dynamics and cell division.

Presumably, this is because of the lower binding afnity of TAXOL® when

conjugated to a uorescent dye. The stains are supplied with a vial of verapamil,

an efux pump inhibitor that may improve probe retention and staining in certain

cell types. ViaFluor® Live Cell Microtubule Stains are not suitable for xation

after staining and cannot be used with xed cells or tissues.

Biotium offers blue uorescent ViaFluor® 405, green uorescent ViaFluor® 488,

and far-red uorescent ViaFluor® 647 Live Cell Microtubule Stains. ViaFluor®

647 Microtubule Stain is compatible with super-resolution imaging by STED.

ViaFluor® Live Cell Microtubule Stains Phalloidin conjugates

Phalloidin is a toxin isolated from the Amanita phalloides mushroom. It is a

bicyclic peptide that binds specically to F-actin (Fig. 2). Unlike antibodies,

the binding of phalloidin is not species-specic, and non-specic staining is

negligible. Biotium offers phalloidin conjugated to a large selection of CF® dyes

which are unrivaled in brightness, photostability, and water solubility compared

to competing dyes such as Alexa Fluor®, DyLight®, and Cy® dyes. A number

of our CF® dyes have also been validated for super-resolution imaging by

STORM, STED, SIM, and other methods.

Cytoskeleton Probes

Cat #. Product Ex/Em (nm)

00049 CF®350 Phalloidin 347/448

00034 CF®405M Phalloidin 408/452

00054 CF®430 Phalloidin 426/498

00055 CF®440 Phalloidin 440/515

00042 CF®488A Phalloidin 490/515

00051 CF®532 Phalloidin 527/558

00043 CF®543 Phalloidin 541/560

00040 CF®555 Phalloidin 555/565

00044 CF®568 Phalloidin 562/583

00045 CF®594 Phalloidin 593/614

00046 CF®633 Phalloidin 630/650

00050 CF®640R Phalloidin 642/662

00041 CF®647 Phalloidin 650/665

00052 CF®660C Phalloidin 667/685

00047 CF®660R Phalloidin 663/682

00053 CF®680 Phalloidin 681/698

00048 CF®680R Phalloidin 680/701

CF Dye® Phalloidin Conjugates

Cat #. Product Ex/Em (nm)

00028 Phalloidin, Biotin-XX N/A

00030 Phalloidin, Fluorescein 496/516

00032 Phalloidin, Rhodamine 110 502/524

00027 Phalloidin, Rhodamine 540/565

00033 Phalloidin, Sulforhodamine 101 (Texas Red®) 591/608

Other Phalloidin Conjugates

www.biotium.com • 11

Endosomal Tracers

Dextrans are water soluble branched-chain polysaccharides. Fluorescently

labeled dextrans are used as markers for trafcking of uid phase endocytic

cargo to lysosomes. In addition, dextrans may be used as tracers for epithelial

and endothelial permeability as well as microinjected tracers for neuronal

morphology. CF® Dye Dextrans are available with a variety of dye colors and

molecular weights ranging from 3,500 to 250,000.

CF® Dye Dextran Conjugates

CF® Dye Transferrin Conjugates

Transferrin is a glycoprotein iron carrier that delivers iron to vertebrate cells

via receptor-mediated endocytosis. Labeled human holo-transferrin is useful

for microscopic studies of the endosomal pathway and/or transferrin uptake.

After transferrin binds to its receptor on the cell surface, it is rapidly internalized

by invagination of clathrin-coated pits. Inside endocytic vesicles, the acidic

environment causes dissociation of iron from the transferrin-receptor complex.

The apotransferrin is then recycled to the plasma membrane. We offer human

transferrin labeled with a selection of our bright and photostable CF® dyes for

imaging of recycling endosomes by microscopy.

CF® Dye Cholera Toxin Subunit B Conjugates

Cholera toxin is the symptom-causing toxin produced by the bacteria Vibrio

cholerae. Cholera toxin subunit B (CT-B) is the non-toxic receptor binding

subunit that binds to ganglioside GM1 in lipid rafts on the plasma membrane.

Fluorescently labeled conjugates of CT-B are useful as endocytic tracers

because the subunit is internalized by clathrin-dependent and -independent

pathways. CT-B conjugates can also be used to label lipid rafts on the cell

surface and as a retrograde neuronal tracer.

Figure 1. Live HeLa cells stained with CF®568 dextran (red)

and counterstained with Hoechst (blue).

Dye Ex/Em (nm) BSA

Cholera

Toxin B

Transferrin

Dextran

3.5K MW

Dextran

10K MW

Dextran

40K MW

Dextran

70K MW

Dextran

150K MW

Dextran

250K MW

CF®350 347/448 --- --- --- 80137 --- --- --- --- ---

CF®488A 490/515 20289 00070 00081 --- 80110 80126 80117 80131 80134

CF®532 527/558 --- 00074 --- --- --- --- --- --- ---

CF®543 541/560 --- 00075 00082 --- 80111 --- --- --- ---

CF®555 555/565 --- --- --- --- 80112 --- --- --- ---

CF®568 562/583 --- 00071 00083 --- 80113 --- --- --- ---

CF®594 593/614 20290 00072 00084 --- 80114 --- --- --- ---

CF®620R 617/639 --- 00076 --- --- --- --- --- --- ---

CF®633 630/650 --- 00077 --- --- --- --- --- --- ---

CF®640R 642/662 20291 00073 00085 --- 80115 --- --- --- ---

CF®660R 663/682 --- 00078 --- --- --- --- --- --- ---

CF®680 681/698 20292 --- --- --- 80118 80127 80129 80132 80135

CF®680R 680/701 --- 00079 00086 --- 80116 --- --- --- ---

CF®750 755/777 --- --- 00087 --- 80119 80128 80130 80133 80136

CF®770 770/797 --- --- 29059 --- 80120 80122 80123 80124 80125

CF®790 784/806 --- --- --- --- 80121 --- --- --- ---

CF ® Dye Labeled Endosomal Tracers

Bovine Serum Albumin CF® Dye Conjugates

Bovine serum albumin (BSA) conjugates are commonly used as protein

tracers for studying the rate of receptor-mediated endocytosis and exocytosis

during vesicle trafcking. Fluorescent BSA conjugates also have been used for

quantitative studies of electroporation and measurement of plasma volume in

rats. We offer BSA conjugates with four of our bright and photostable CF® dyes.

Figure 2. Live HeLa cells were stained with CF®488A

transferrin (green) and DAPI (blue).

v2.27.20

www.biotium.com

General Inquiries: [email protected]

Technical Support: [email protected]

Phone: 800-304-5357

Biotium, Inc.

46117 Landing Parkway

Fremont, CA 94538 USA

About Us

At Biotium, we are dedicated to developing cutting-edge uorescent

solutions for life science research. Our efforts have resulted in a growing

number of unique and industry-leading uorescence-based technologies for

a wide array of biological research applications. Our products are available

in the U.S. through our website, and worldwide through our extensive

network of domestic and international distributors.

We license our technologies to a number of international biotechnology

companies, and collaborate with academic laboratories to develop tools for

the constantly evolving research community. We welcome inquiries about

licensing the use of our dyes, technologies, or trademarks; email us at

[email protected].

Biotium implements a Quality System, certied by QAS according to

Standard QAS ISO 9001:2015.