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Fluo-3

Fluo-3 is a fluorescent calcium indicator used to measure intracellular calcium concentrations.
It is a derivative of the fluorescein dye and exhibits a large fluorescence increase upon binding to calcium.
Fluo-3 has been widely utilized in cellular and physiological research to visualize and quantify calcium dynamics within living cells.
Its high sensitivity and responsiveness to calcium make it a valuable tool for studying calcium-mediated signaling pathways and processes, such as neurotransmitter release, muscle contraction, and apoptosis.
Fluo-3 can be loaded into cells through a variety of methods and its fluorescence can be detected using fluorescence microscopy or flow cytometry.
Reasearchers can leverge PubCompare.ai's AI-powered platform to optimiize their Fluo-3 research by identifying the most accurate and reproducible protocols from literature, pre-prints, and patents to improve their experiments and findings.

Most cited protocols related to «Fluo-3»

Measuring Cytosolic Calcium in Cardiac Progenitor Cells

Cited 13 times

hCPCs were isolated from myocardial specimens obtained from patients who underwent cardiac surgery. Cytosolic Ca²⁺ levels in cultured hCPCs were measured utilizing the Ca²⁺ indicator Fluo-3 and two-photon microscopy. Cell proliferation in vivo and in vitro was evaluated by BrdUrd incorporation. Results are shown as mean±SEM. An expanded Materials and Methods section can be found in the online data supplement available at http://circres.ahajournals.org.

Ferreira-Martins J., Rondon-Clavo C., Tugal D., Korn J.A., Rizzi R., Padin-Iruegas M.E., Ottolenghi S., De Angelis A., Urbanek K., Iwata N., D’Amario D., Hosoda T., Leri A., Kajstura J., Anversa P, & Rota M. (2009). Spontaneous Calcium Oscillations Regulate Human Cardiac Progenitor Cell Growth. Circulation research, 105(8), 764-774.

Publication 2009

Cell Proliferation Cytosol Dietary Supplements Fluo-3 Microscopy Myocardium Patients Surgical Procedure, Cardiac

Functional Imaging of Mitochondrial Dynamics

Cited 12 times

Functional time-lapse recordings of mitochondria were mostly performed with a fluorescence imaging system composed of a Xenon light source (Polychrome II, Till Photonics) and a sensitive CCD camera (Imago QE, 62% quantum efficiency at 500 nm, PCO Imaging) attached to an upright microscope (Axiotech Vario, Zeiss). JC-1 was excited at 490 nm and an optical image splitter device (Dual-View^TM, Optical Insights) was attached to the microscope to separate spectrally the green and red components of JC-1 fluorescence (see Fig. 2d for schematic and filter settings). For fluo 3-AM recordings a 505 nm beamsplitter, a 535/35 nm bandpass filter (emitter) and 485-nm excitation wavelength were used. Experiments were performed in a submersion style chamber; cell cultures were continuously superfused with ACSF (32–33°C, flow rate 3–4 ml/min) and 63× or 100× water immersion objectives (Apochromat and Achroplan, respectively; Zeiss) were used.
High-resolution ratiometric JC-1 analyses were performed using a custom-built two-photon laser scanning microscope (TPLSM) [29 ]. The original system layout has been extended by a second upright microscope (BX51WI, Olympus) equipped with an IR-optimized 20× 0.95NA objective (XLUMPlanFL, Olympus) and two non-descanned single-photon counting photomultiplier tubes (H7421-40, Hamamatsu; Fig. 2a). Also a new laser (Mai Tai eHP DS, Newport Spectra-Physics) has been added just recently. The general scan head design was unchanged, but improved and faster galvanometric scanners (VM500C with MiniSAX control circuits and 4 mm mirrors; General Scanning/Cambridge Technology) were chosen. The scanning process and data acquisition were controlled by a digital signal processor (ADwin-Gold-ENET; Jäger) and the outputs of the two photomultiplier tubes (TTL pulses) were analyzed by a custom-built 2-channel TTL pulse counter. Z-axis (axial) adjustment was realized by a piezo-driven objective nano-positioning system (Pifoc P-721; Physik Instrumente). For two-photon imaging, the excitation wavelength was 790–800 nm; fluorescence was separated using a 670-nm dichroic mirror (670dcxxr) and the green and red components were further separated by a 570-nm dichroic mirror (570dcxr) followed by 536/40 and 617/73 nm bandpass emission filters, respectively. Offline image analysis was performed with Tillvision 4.0 (Till Photonics) and MetaMorph Offline 6.1/7.0 (Molecular Devices).

Keil V.C., Funke F., Zeug A., Schild D, & Müller M. (2011). Ratiometric high-resolution imaging of JC-1 fluorescence reveals the subcellular heterogeneity of astrocytic mitochondria. Pflugers Archiv, 462(5), 693-708.

Publication 2011

Cell Culture Techniques Epistropheus Fingers Fluo-3 Fluorescence Gold Head Laser Scanning Microscopy Light Medical Devices Microscopy Mitochondria Pulse Rate Pulses Submersion Xenon

Fluorescent Assays for Cellular Stress

Cited 10 times

The determination of calcium accumulation, MMP disruption, and ROS generation was reported previously [24 (link),65 (link)]. Fluorescent calcium indicator (Fluo 3, 5 mM), JC-1 cationic dye (1.25 μg/mL), and the carboxy derivative of fluorescein (carboxy-H2DCFDA, 1.0 mM) were used to determine calcium accumulation, MMP disruption, and ROS generation, respectively. A specific fluorescent dye was used to label cells were labeled for 30 min after treating the cells with different concentrations of 13-AC. PBS was used for washing. Flow cytometry was used to determine the changes of ROS, MMP, and calcium concentrations.

Liu Y.C., Peng B.R., Hsu K.C., El-Shazly M., Shih S.P., Lin T.E., Kuo F.W., Chou Y.C., Lin H.Y, & Lu M.C. (2020). 13-Acetoxysarcocrassolide Exhibits Cytotoxic Activity against Oral Cancer Cells through the Interruption of the Keap1/Nrf2/p62/SQSTM1 Pathway: The Need to Move Beyond Classical Concepts. Marine Drugs, 18(8), 382.

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Publication 2020

2',7'-dichlorodihydrofluorescein diacetate Calcium Cations Flow Cytometry Fluo-3 Fluorescein Fluorescent Dyes

Quantifying Cardiomyocyte Calcium Dynamics

Cited 8 times

[Ca²⁺]_i was quantified with Fluo-3-acetoxymethyl (Fluo-3) ester in bath and pipette solution. After de-esterification, fluorescence was excited at 488 nm and emitted light (>520 nm) converted to [Ca²⁺]_i assuming

{[{Ca}^{2 +}]}_{i} = k_{d} (\frac{F}{F_{\max} - F})

where k_d is the dissociation constant of Fluo-3 (864 nmol/L), F=Fluo-3 fluorescence, and F_max is Ca²⁺-saturated fluorescence obtained at the end of each experiment.¹⁷Membrane-currents and APs were recorded at 37°C in whole-cell ruptured-patch configuration using voltage/current-clamp techniques with simultaneous [Ca²⁺]_i measurement. There was no significant difference in membrane capacitance between pAF (102.0±11.7 pF, n=15/9 [myocytes/patients]) and Ctl (113.6±6.1 pF, n=35/25; P=0.340) myocytes. Currents are expressed as current-densities (pA/pF). L-type Ca²⁺-current (I_Ca,L)-triggered [Ca²⁺]_i-transients were recorded simultaneously, as previously described.^{15 (link)} Sarcoplasmic-reticulum (SR) Ca²⁺-leak was measured as the decrease in [Ca²⁺]_i following application of tetracaine in the absence of extracellular Ca²⁺/Na⁺, as described by Shannon et al.^{18 (link)}

Voigt N., Heijman J., Wang Q., Chiang D.Y., Li N., Karck M., Wehrens X.H., Nattel S, & Dobrev D. (2013). Cellular and Molecular Mechanisms of Atrial Arrhythmogenesis in Patients with Paroxysmal Atrial Fibrillation. Circulation, 129(2), 145-156.

Publication 2013

Bath Cells Esterification Esters Fluo-3 Fluorescence Light Muscle Cells Patients Sarcoplasmic Reticulum Tetracaine Tissue, Membrane Transients

Characterization of Punctate Fluorescence Hot Spots

Cited 8 times

Most images recorded in this study were x-t scans. Toward the end of some experiments, however, occasional x-y scans were also recorded. An unusual feature of some of these scans was the presence of punctate regions of high fluorescence (“hot spots”; Fig. 2 A). Hot spots do not appear to be caused by transient elevations in myoplasmic [Ca²⁺] because their appearance was unchanged in a second x-y scan of the same fiber location (hence hot spots reflect persistent rather than transient events), and their center location was always ∼0.5 μm from the z-line (whereas sparks are centered at the z-line; see results). The physical basis of hot spots is unknown, but a likely possibility is that they arise from a population of fluo-3 molecules that are sequestered inside a small organelle with elevated free [Ca²⁺].
In x-t images, hot spots appeared as bright streaks (“streakers”) that were offset ∼0.5 μm from a z-line. Interestingly, streakers, when detected, were always present at the beginning of an x-t image and usually faded during the course of the image, probably because the responsible fluo-3 molecules were bleached by the high intensity light that repeatedly scanned the same spatial location. This might occur if fluo-3 molecules in the Ca²⁺-bound form are more readily bleached than those in the Ca²⁺-free form or if bleached fluo-3 cannot be replenished by diffusion. Fig. 2 B shows an example of a fading streaker; the narrow band of high fluorescence at the start of the image faded to background levels over a period of ∼1 s. These data are shown in F units (counts per microsecond, proportional to fluorescence intensity; see scale at right). Fig. 2 C shows a portion of Fig. 2 B after expansion of the x- and t-axes and normalization by resting F (ΔF/F units). In Fig. 2 D, the spatial profile of the streaker has been fitted with a Gaussian function () to estimate FWHM_X (); the fitted value is 0.27 μm. In eight similar streakers from five fibers, the value of FWHM_X was 0.30 ± 0.01 μm (mean ± SEM). Other streakers, which had somewhat larger values of FWHM_X, were not included in this analysis because of the possibility that they were out of focus. The value 0.30 μm represents an upper limit of the in vivo FWHM_X of the PSF of the microscope. This value is ∼0.1 μm larger than the value of FWHM_X estimated with 0.1 μm fluorescent beads (0.21 ± 0.01 μm; see above), probably because of the nonzero size of the organelle or other structure(s) that contains the fluo-3 molecules responsible for a streaker.

Hollingworth S., Peet J., Chandler W.K, & Baylor S.M. (2001). Calcium Sparks in Intact Skeletal Muscle Fibers of the Frog. The Journal of General Physiology, 118(6), 653-678.

Publication 2001

Diffusion Epistropheus Exanthema Fibrosis Fluo-3 Fluorescence Intravital Microscopy Light Organelles Physical Examination Transients

Most recents protocols related to «Fluo-3»

Cytosolic Free Ca2+ Quantification

To measure relative levels of cytosolic free Ca²⁺, cells were stained with 3 mmol/L Fluo-4-AM for 25 min, washed twice with Dulbecco phosphate-buffered saline (DPBS), resuspended in DPBS without Ca²⁺, Mg²⁺ (if not otherwise indicated) plus 20 mmol/L HEPES and maintained at 37C while treating the cells as indicated and analyzing by BD FACSVerse flow cytometer (FL-1 channel).

Liu B., Chen R., Zhang Y., Huang J., Luo Y., Rosthøj S., Zhao C, & Jäättelä M. (2023). Cationic amphiphilic antihistamines inhibit STAT3 via Ca2+-dependent lysosomal H+ efflux. Cell Reports, 42(2), 112137.

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Publication 2023

Cells Cytosol Fluo-3 HEPES Phosphates Saline Solution

Preparation and Treatment of HEK Cells

U73122 (S8011, Selleck Chemicals, Houston, TX, USA) is dissolved in dimethyl sulfoxide (DMSO, D8371, Beijing, China), with the final concentration of 9 mM, and stored at −20 °C. For subsequent experiments, combined with cell viability results (see Supplementary data: Supplementary Fig s9a) and previous research results^{49 (link),54 (link)}, we selected 9 μM U73122 treatment of HEK for 15 min.
MK-2206 (2HCl) (S1078, Houston, TX, USA) is dissolved in dimethyl sulfoxide (DMSO, D8371, Beijing, China), with the final concentration of 12 mM, and stored at −80 °C. For subsequent experiments, combined with cell viability results (see Supplementary data: Supplementary Fig s9a) and previous research results^{55 (link)}, we selected 12 μM MK-2206 treatment of HEK for 24 h.
Pertussis toxin (PTX, P7208, Sigma-Aldrich St., Louis, USA) is dissolved in ddH₂O, with the final concentration of 200 μg/ml, and stored at 4 °C. For subsequent experiments, combined with cell viability results (see Supplementary data: Supplementary Fig s9a) and previous research results^{48 (link)}, we selected 200 ng PTX treatment of HEK for 4 h.
Fluo-3 AM (F1241, Invitrogen, Thermo Fisher Scientific, USA) is dissolved in dimethyl sulfoxide (DMSO, D8371, Beijing, China), with the final concentration of 3 mM and stored at the light-free conditions at −20 °C.

Lan Y., Zeng W., Wang Y., Dong X., Shen X., Gu Y., Zhang W, & Lu H. (2023). Opsin 3 mediates UVA-induced keratinocyte supranuclear melanin cap formation. Communications Biology, 6, 238.

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Publication 2023

Cell Survival Fluo-3 Light MK 2206 Pertussis Toxin Sulfoxide, Dimethyl U 73122

Calcium Imaging and Flux Assay

For calcium imaging experiments, cells were collected and incubated with 3 μM Fluo-3 AM for 15 min at 37 °C in the darkroom. Fluorescent images of Fluo-3AM-loaded cells were acquired using Cell Observer-Living Cells (Zeiss, German).
For calcium flux experiments, Fluo-3AM-loaded cells were centrifuged at 1000 rpm for 5 min at RT, washed once with PBS, and resuspended with 500 μl PBS. The intracellular calcium concentration was detected by flow cytometry (BD Biosciences, San Jose, CA, USA). The excitation source for Fluo-3 AM was a 488-nm air-cooled argon laser, and the emission was measured using a 525-nm band-pass filter.

Lan Y., Zeng W., Wang Y., Dong X., Shen X., Gu Y., Zhang W, & Lu H. (2023). Opsin 3 mediates UVA-induced keratinocyte supranuclear melanin cap formation. Communications Biology, 6, 238.

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Publication 2023

Argon Ion Lasers Calcium Cells Flow Cytometry Fluo-3 FLUOS Protoplasm

Intracellular Ca2+ Levels in MCF-7-ADR Cells

To investigate the effects of TPGS on intracellular Ca²⁺ levels, MCF-7-ADR cells were inoculated into 6-well plates (1 × 10⁶ cells/well) and cultured in medium containing different TPGS concentrations (16, 80, and 400 µg/mL) for 48 h. Cells were then centrifuged at 8000 rpm at 4 °C for 5 min and washed twice in cold PBS. Next, the fluorescent fluo-3 AM probe was diluted in PBS (1:1000), added to cells, and incubated at 37 °C for 0.5 h before washing twice in PBS. Fluorescence at 528 nm was determined by flow cytometry in the FL1 channel.

Tang L., Huang K., Jiang W., Fu L., Zhang R., Shen L., Ou Z., Huang Y, & Zhang Z. (2023). Exploration of the inhibition action of TPGS on tumor cells and its combined use with chemotherapy drugs. Drug Delivery, 30(1), 2183830.

Publication 2023

Cells Cold Temperature Flow Cytometry Fluo-3 Fluorescence MCF-7 Cells Protoplasm tocophersolan

Monitoring Amyloid-Beta Induced Calcium Dynamics

2 × 10⁵ HT22 cells were incubated in 35 mm confocal dishes for 24 h at 37 °C in a CO² incubator. A 5 mM Fluo-3 AM stock solution was diluted with Hanks-Balanced Salt Solution (HBSS) to a 5 μM working solution and then added to the cells for 30 min at 37 °C. HT22 cells were washed three times with HEPES-buffered saline and then incubated in an imaging chamber at 37 °C for an additional 10 min. After the addition of 30 μM of Aβ aggregates in HBSS buffer, changes in cellular Ca²⁺ levels were monitored by epifluorescence microscopy (Applied Precision DeltaVision Elite, Applied Precision, Inc., United States) using the real-time mode for tracking Fluo-3 changes for 5 min. The data inspection program provided by DeltaVision software was used to measure the intensity of Fluo-3 fluorescence.

Song L.L., Qu Y.Q., Tang Y.P., Chen X., Lo H.H., Qu L.Q., Yun Y.X., Wong V.K., Zhang R.L., Wang H.M., Liu M.H., Zhang W., Zhang H.X., Chan J.T., Wang C.R., Wu J.H, & Law B.Y. (2023). Hyperoside alleviates toxicity of β-amyloid via endoplasmic reticulum-mitochondrial calcium signal transduction cascade in APP/PS1 double transgenic Alzheimer's disease mice. Redox Biology, 61, 102637.

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Publication 2023

Buffers Cells Fluo-3 Fluorescence Hanks Balanced Salt Solution HEPES Hyperostosis, Diffuse Idiopathic Skeletal Microscopy Saline Solution

Top products related to «Fluo-3»

Fluo 3 am by Thermo Fisher Scientific

Sourced in United States, China, United Kingdom, Germany, Italy, Austria

Fluo-3 AM is a fluorescent calcium indicator used to measure intracellular calcium levels in cells. It binds to calcium ions and emits fluorescence upon excitation, allowing researchers to monitor calcium dynamics within cellular environments.

Fluo 3 am by Beyotime

Sourced in China, United States

Fluo-3 AM is a fluorescent calcium indicator used for the detection and measurement of intracellular calcium levels. It is a cell-permeable, acetoxymethyl (AM) ester derivative of the fluorescent dye Fluo-3.

Fluo 3 am by Merck Group

Sourced in United States, Germany, Sao Tome and Principe, Belgium

Fluo-3/AM is a fluorescent calcium indicator used for the detection and measurement of intracellular calcium levels. It is a cell-permeant acetoxymethyl (AM) ester derivative of the fluorescent dye Fluo-3.

Fluo 3 by Thermo Fisher Scientific

Sourced in United States

Fluo-3 is a fluorescent calcium indicator used in biological research. It binds to intracellular calcium ions, allowing researchers to monitor calcium levels and dynamics within cells.

Fluo 3 am by Dojindo Laboratories

Sourced in Japan, China

Fluo 3-AM is a fluorescent calcium indicator used for monitoring intracellular calcium levels. It is a cell-permeable form of the Fluo-3 dye that can be readily loaded into cells. Fluo 3-AM allows for the detection and quantification of changes in intracellular calcium concentration.

Pluronic f 127 by Thermo Fisher Scientific

Sourced in United States, Germany, United Kingdom, Japan, Belgium, Switzerland, Italy, Lithuania, Canada, India

Pluronic F-127 is a non-ionic, surfactant-based material commonly used in various laboratory applications. It is a triblock copolymer composed of polyethylene oxide and polypropylene oxide segments. Pluronic F-127 is known for its ability to form thermoreversible gels and has versatile applications in areas such as drug delivery, tissue engineering, and cell culture.

Facscalibur by BD

Sourced in United States, Germany, United Kingdom, China, Canada, Japan, Italy, France, Belgium, Switzerland, Singapore, Uruguay, Australia, Spain, Poland, India, Austria, Denmark, Netherlands, Jersey, Finland, Sweden

The FACSCalibur is a flow cytometry system designed for multi-parameter analysis of cells and other particles. It features a blue (488 nm) and a red (635 nm) laser for excitation of fluorescent dyes. The instrument is capable of detecting forward scatter, side scatter, and up to four fluorescent parameters simultaneously.

Fetal bovine serum (fbs) by Thermo Fisher Scientific

Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal

Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

Pluronic f 127 by Merck Group

Sourced in United States, Germany, United Kingdom, China, Italy, India, Spain, Switzerland, Sao Tome and Principe, France, Japan, Australia, Singapore, Macao, Brazil, Denmark, Austria

Pluronic F-127 is a non-ionic, tri-block copolymer composed of polyethylene oxide (PEO) and polypropylene oxide (PPO) segments. It is a widely used surfactant and emulsifying agent in various pharmaceutical and biomedical applications.

Rhod 2 am by Thermo Fisher Scientific

Sourced in United States, Germany, United Kingdom, Canada

Rhod-2 AM is a fluorescent calcium indicator. It is used for the detection and measurement of intracellular calcium levels in live cells.

What are some common challenges researchers face when using Fluo-3 for intracellular calcium measurements?

One of the main challenges with Fluo-3 is ensuring accurate and reliable calcium measurements. Factors like dye loading efficiency, compartmentalization, and leakage can influence the fluorescence signal and lead to inconsistent results. Researchers must optimize staining protocols and consider potential interference from other ions or cellular processes. Additionally, the high sensitivity of Fluo-3 can make it susceptible to photobleaching, requiring careful experimental design and imaging conditions.

How can PubCompare.ai help researchers optimize their Fluo-3 experiments?

PubCompare.ai's AI-powered platform can be incredibly helpful for researchers using Fluo-3. The platform allows you to efficiently screen protocol literature from publications, preprints, and patents to identify the most effective and reproducible Fluo-3 staining and imaging methods. The AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for your specific research goals and improve the accuracy and reliability of your calcium measurements. PubCompare.ai can also help you compare different Fluo-3 dyes and formulations to find the optimal product for your experiments.

What are some of the different variations or types of Fluo-3 available for researchers?

Fluo-3 comes in several different formulations and derivatives to suit various experimental needs. There are cell-permeable acetoxymethyl (AM) ester versions of Fluo-3 that can be loaded into cells, as well as salt forms that are better suited for in vitro or microfluidic applications. Researchers can also find Fluo-3 conjugated to dextran or other carriers to improve dye retention and reduce leakage. Additionally, there are Fluo-3-based calcium indicators with different calcium binding affinities and spectral properties, such as Fluo-4 and Fluo-5N, which can be selected based on the expected calcium concentrations in the system of interest.

What are some key applications of Fluo-3 in cellular and physiological research?

Fluo-3 has been widely used to study a variety of calcium-mediated processes in cells and tissues. It is commonly employed to visualize and quantify calcium signaling events, such as neurotransmitter release, muscle contraction, and apoptosis. Fluo-3 has also been utilized to investigate calcium dynamics in excitable cells like neurons and cardiomyocytes, as well as in non-excitable cells like immune cells and stem cells. Additionally, Fluo-3 has been used to monitor calcium-dependent enzymes activities and to track calcium changes during cell differentiation and development. The versatility of Fluo-3 makes it a valuable tool for a broad range of cellular and physiological research applications.

How can researchers ensure they are using the most effective and reproducible Fluo-3 protocols?

Ensuring the use of effective and reproducible Fluo-3 protocols is crucial for obtaining accurate and reliable calcium measurements. This is where PubCompare.ai can be particularly helpful. The platform's AI-powered analysis can sceen protocol literature from publications, preprints, and patents to identify the most optimal Fluo-3 staining and imaging methods. By leveraging PubCompare.ai, researchers can compare different protocols, identify key differences in their effectiveness, and select the best approach for their specific research goals. This can help improve the reproducibility and accuracy of their Fluo-3-based experiments and lead to more meaningful scientific findings.

More about "Fluo-3"

Fluo-3, Fluo-3 AM, Fluo-3/AM, Fluo 3-AM, Pluronic F-127, FACSCalibur, FBS, Rhod-2 AM