VECTASHIELD. Mounting Medium PRODUCT CATALOG

Transcription

1 VECTASHIELD Mounting Medium PRODUCT CATALOG

2 VECTASHIELD Mounting Media are unsurpassed in preventing photobleaching. The different formulations of VECTASHIELD Mounting Media all offer the same outstanding anti-fade and antiphotobleaching properties. They are all compatible with fluorescein, Texas Red, AMCA, DyLight dyes, Alexa Fluor dyes, fluorescent nuclear stains, fluorescent proteins, fluorescent tracers, histochemical stains, and most fluorochromes. Features of VECTASHIELD Mounting Media: Inhibit photobleaching of most fluorochromes Available in non-hardening or hardening formulations Available with or without DAPI or propidium iodide No warming necessary Slides can be viewed after prolonged storage Continues to inhibit photobleaching after prolonged storage of mounted slides Easy to use Optically clear Ideal refractive index The original VECTASHIELD Mounting Medium is a glycerol-based, aqueous mountant that does not solidify but remains a viscous liquid on the slide. After mounting, coverslipped slides will not readily dry out and can be reviewed for weeks afterwards without sealing. For prolonged storage, coverslips can be permanently sealed around the perimeter with nail polish. Mounted slides should be stored at 4 ºC. Product Counterstain Cat. No. Unit Size Hardening Refractive Index VECTASHIELD Mounting Medium None H ml No 1.45 DAPI H ml No 1.45 PI H ml No 1.45 VECTASHIELD Hard Set Mounting Medium None H ml Yes 1.46 DAPI H ml Yes 1.46 TRITC-Phalloidin H ml yes 1.46 VECTASHIELD Mounting Media are compatible with a wide array of fluorochromes, enzymatic substrates, and fluorescent proteins. Please consult the compatibility table (located in product information box) to determine if VECTASHIELD will be compatible in your system. VECTASHIELD Mounting Medium Antifade Comparison Other manufacturers measure the antifade properties of their mountants using labeled microspheres or arrayed spots. Vector Labs prefers to measure antifade properties of VECTASHIELD mountants using frozen tissue sections immunohistochemically stained with fluorescently labeled secondary antibodies. Antifade capability is measured using a 40x objective with real time imaging over 30 seconds of continuous exposure to the excitation illumination. Individual intensity measurements are recorded from 6 separate labeled regions and the average is calculated. The intensity after 30 second exposure is expressed as a percentage of the intensity at zero time. The values for PG are taken from the manufacturer s published results.

3 VECTASHIELD Mounting Medium Antifade Properties Fluorescent Dye Remaining Fluorescence (%) DyLight % Alexa Fluor % Fluorescein 98% Alexa Fluor % DyLight % Tetramethylrhodamine 97% Cy 3 94% DyLight % Alexa Fluor % Texas Red 100% DyLight % Antifade properties are measured using frozen tissue sections immunochistochemically stained with fluorescently labeled secondary antibodies. Antifade capability is measured using a 40x objective with real time imaging over 30 sec of continuous exposure to the excitation illumination. Individual intensity measurements are recorded from 6 separate labeled regions and the average is calculated. The intensity after 30 sec exposure is expressed as a percentage of the intensity at zero time. Absence pf a particular fluorochrome does not indicated incompatibility. VECTASHIELD Mounting Medium in Super Resolution Microscopy The optimal medium for super-resolution imaging methods maximizes the lifetime and photo-switching characteristics of fluorophores. VECTASHIELD Mounting Medium has been shown to be compatible with a number of fluorophores used in super resolution methods including STORM, STED, and 3D-SIMM imaging, such as Cy5 or Alexa fluor 647, resulting in greater convenience and reproducibility of the method. (Olivier, N. et al., 2013, Biomedical Optics Express, Vol. 4 No. 6, pp ). Please note: VECTASHIELD Mounting Medium may not be compatible with all enzymatic substrates or fluorescent proteins and test sections would be advisable when using anything other than commonly used fluorochromes. The refractive index for VECTASHIELD Mounting Medium is R.J. Florijn, et. al. Cytometry, 19 (1995)

4 VECTASHIELD Mounting Medium Compatibility Based on in-house testing and data from published papers, Vectashield Mounting Medium has been used with the following fluorochromes. Absence of a particular fluorochrome does not indicate incompatibility. 1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine 1,1'-dioctadecyl-3,3,3',3'- tetramethylindodicarbocyanine calceincalcofluor white Cascade Blue FluoProbes 547 fluorescein fluorescent brightener 28 Rhodamine Green Rhodamine Red SpectrumAqua 3,3'-dioctadecyloxacarbocyanine CellTracker Blue Fluoro-Emerald SpectrumBlue 4-(4-(dihexadecylamino)styryl)- N-methylpyridinium 4-chloromethyl benzoyl amino tetramethyl rhodamine CellTracker Green CellTracker Orange CellTracker Red Fluoro-Gold Fluoro-Jade Fluoro-Ruby SpectrumGold SpectrumGreen SpectrumOrange 5-(4,6-dichlorotriazinyl) aminofluorescein chromomycin a3 coumarin- green fluorescent protein hoechst SpectrumRed SYBR Gold 5-chloromethylfluorescein diacetate 7-diethylaminocoumarin-3- carboxylic acid Cy 2 Cy 3 Cy 3.5 hoechst lissamine rhodaminelucifer yellow SYBR Green SYTO SYTOX Green 7-nitrobenz-2-oxa-1,3-diazole Cy 5 LysoTracker SYTOX Orange acridine orange Cy 5.5 LysoTracker Red tamra- actinomycin d Cy 7 Marina Blue tetramethylrhodamine Alexa Fluor 350 DAPI MitoTracker Texas Red Alexa Fluor 405 Dead Red MitoTracker Deep Red thioflavin s Alexa Fluor 488 dihydroethidium MitoTracker Green TO-PRO Alexa Fluor 532 distamycin a MitoTracker Orange TO-PRO -1 Alexa Fluor 546 doxorubicin MitoTracker Red TO-PRO -3 Alexa Fluor 555 DRAQ5 NeuroTrace TOTO -1 Alexa Fluor 568 Dy547- nile red TOTO -3 Alexa Fluor 594 DyLight 488 nonyl acridine orange Vector Red Substrate Kit Alexa Fluor 633 DyLight 549 NothernLights -557 yellow fluorescent protein Alexa Fluor 647 DyLight 594 oil red o YO-PRO -1 Alexa Fluor 660 DyLight 649 Oregon Green 488 YOYO Alexa Fluor 680 ER-Tracker Blue/white Oregon Green 514 YOYO -1 alizarin red s ER-Tracker Red Pacific Blue allophycocyanin ethidium PicoGreen AMCA ethidium bromide propidium iodide aminoactinomycin d ethidium homodimer quantum dot- bisbenzimide evans blue red fluorescent protein blue fluorescent protein fast blue rhodamine BODIPY FluoProbes 488 rhodamine 600 Trademarks are the properties of the respective companies.

5 PRODUCT INFORMATION Product VECTASHIELD Mounting Medium Catalog No. H-1000 Amount 10 ml Storage Conditions Refrigerate in the dark VECTASHIELD Mounting Medium is a unique, stable formula for preserving fluorescence. VECTASHIELD Mounting Medium prevents rapid photobleaching of fluorescent proteins and fluorescent dyes (see website for compatibility). VECTASHIELD Mounting Medium is supplied in a bottle fitted with a screw cap. A cap fitted with a drop dispenser pipet is also included. The screw cap can be replaced with the drop dispenser cap for routine use and during storage. Once the drop dispenser cap has been secured on the bottle, the bottle should be stored in an upright position. To mount tissues or cells on a slide, dispense VECTASHIELD Mounting Medium onto the specimen. The pipet supplied with the product is designed to deliver small drop volumes of approximately 25 µl. Coverslip and allow VECTASHIELD Mounting Medium to disperse over the entire section. VECTASHIELD Mounting Medium does not solidify, but remains a liquid on the slide and can be stored without sealing. If desired, coverslips can be sealed around the perimeter with nail polish or a plastic sealant. Mounted slides should be stored at 4 C, protected from light. VECTASHIELD Mounting Media Products: Product Counterstain Cat. No. Unit Size Hardening Refractive Index* VECTASHIELD Mounting Medium none H ml no 1.45 DAPI H ml no 1.45 PI H ml no 1.45 VECTASHIELD HardFSet Mounting Medium none H ml yes 1.46 DAPI H ml yes 1.46 TRITC-Phalloidin H ml yes 1.46 *Measured at room temperature. VECTASHIELD Hard F Set Mounting Medium measured after hardening. VECTASHIELD is a registered trademark of Vector Laboratories, Inc. Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA U.S.A. Tel: (650) Fax: (650) vector@vectorlabs.com Web site: 01/15

6 Product Catalog No. H-1200 Amount 10 ml Concentration of DAPI Storage Conditions PRODUCT INFORMATION VECTASHIELD Mounting Medium with DAPI 1.5 µg/ml Refrigerate in the dark VECTASHIELD Mounting Medium is a unique, stable formula for preserving fluorescence. VECTASHIELD Mounting Medium prevents rapid photobleaching of fluorescent proteins and fluorescent dyes (see website for compatibility). This special formulation of VECTASHIELD Mounting Medium contains 4,6-diamidino-2-phenylindole (DAPI), which fluoresces when bound to DNA and is used as a nuclear or chromosomal counterstain. This product is designed to be used for in situ hybridization techniques or other procedures requiring fluorescent labeling of DNA. DAPI may also stain RNA but displays greater selectivity for binding DNA over RNA. DAPI excites at about 360 nm and emits at about 460 nm when bound to DNA, producing a blue fluorescence. VECTASHIELD Mounting Medium is supplied in a bottle fitted with a screw cap. A cap fitted with a drop dispenser pipet is also included. The screw cap can be replaced with the drop dispenser cap for routine use and during storage. Once the drop dispenser cap has been secured on the bottle, the bottle should be stored in an upright position. To mount tissues or cells on a slide, dispense VECTASHIELD Mounting Medium onto the specimen. The special pipet supplied with the product is designed to deliver small drop volumes of approximately 25 µl. Coverslip and allow VECTASHIELD Mounting Medium to disperse over the entire specimen. VECTASHIELD Mounting Medium does not solidify, but remains a liquid on the slide and can be stored without sealing. If desired, coverslips can be permanently sealed around the perimeter with nail polish or a plastic sealant. Mounted slides should be stored at 4 C protected from light. Note: DAPI is a mutagen and appropriate precautions should be taken when using it. VECTASHIELD Mounting Media Products: Product Counterstain Cat. No. Unit Size Hardening Refractive Index* VECTASHIELD Mounting Medium none H ml no 1.45 DAPI H ml no 1.45 PI H ml no 1.45 VECTASHIELD HardFSet Mounting Medium none H ml yes 1.46 DAPI H ml yes 1.46 TRITC-Phalloidin H ml yes 1.46 *Measured at room temperature. VECTASHIELD Hard F Set Mounting Medium measured after hardening. VECTASHIELD is a registered trademark of Vector Laboratories, Inc. 01/15 Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA U.S.A. Tel: (650) Fax: (650) vector@vectorlabs.com Web site:

7 VECTASHIELD Hard Set Mounting Medium Compatibility Based on in-house testing and data from published papers, Vectashield Hard Set Mounting Medium has been used with the following fluorochromes. Absence of a particular fluorochrome does not indicate incompatibility. 1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine Alexa Fluor 350 Alexa Fluor 488 Alexa Fluor 546 Alexa Fluor 555 Alexa Fluor 568 Alexa Fluor 594 Alexa Fluor 633 Alexa Fluor 635 Alexa Fluor 647 AMCA Cy 2 Cy 3 Cy 5 DAPI DyLight 488 DyLight 549 DyLight 594 DyLight 649 fluorescein green fluorescent protein hoechst dye phycoerythrin propidium iodide rhodamine red fluorescent protein SpectrumGreen SpectrumOrange SpectrumRed SYTOX Green tetramethylrhodamine Texas Red TO-PRO -3 Trademarks are the properties of the respective companies.

8 PRODUCT INFORMATION Product VECTASHIELD Hard F Set Mounting Medium Catalog No. H-1400 Amount 10 ml Storage Conditions Refrigerate in the dark VECTASHIELD Hard F Set Mounting Medium preserves fluorescence and hardens after coverslipping. VECTASHIELD Hard F Set Mounting Medium s unique, stable formula prevents rapid photobleaching of fluorescent proteins and fluorescent dyes (see website for compatibility). VECTASHIELD Hard F Set Mounting Medium is supplied in a bottle fitted with a screw cap. A cap fitted with a drop dispenser pipet is also included. The screw cap can be replaced with the drop dispenser cap for routine use and during storage. Once the drop dispenser cap has been secured on the bottle, the bottle be should be stored in an upright position. To mount tissues or cells on a slide, dispense VECTASHIELD Hard F Set Mounting Medium onto the specimen. The pipet supplied with the product is designed to deliver small drop volumes of approximately 25 µl. Coverslip and allow VECTASHIELD Hard F Set Mounting Medium to disperse over the entire specimen. After approximately 15 minutes at room temperature, the coverslip will become immobilized, and optimal antifade ability and refractive index will be achieved. After curing at room temperature for 15 minutes, slides can be placed at 4 ºC, and the mounting media will harden completely overnight. Mounted slides should be stored at 4 C or -20 ºC, protected from light. For prolonged storage, -20 ºC is recommended. If retraction occurs during prolonged storage, remove coverslip and remount. Coverslips can be easily removed by soaking in PBS overnight. Notes: When mounting thick sections, adjustments to the mounting protocol may be required. Use of non-hardening VECTASHIELD Mounting Medium (H-1000) may be preferred. The nuclear counterstains, DAPI and PI, will diffuse out of nuclei over time if the cells have been fixed in 100% methanol. In this case non-hardening VECTASHIELD Mounting Medium may be preferred. (over) Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA U.S.A. Tel: (650) Fax: (650) vector@vectorlabs.com Web site:

9 VECTASHIELD Mounting Media Products: Product Counterstain Cat. No. Unit Size Hardening Refractive Index* VECTASHIELD Mounting Medium none H ml no 1.45 DAPI H ml no 1.45 PI H ml no 1.45 VECTASHIELD HardFSet Mounting Medium none H ml yes 1.46 DAPI H ml yes 1.46 TRITC-Phalloidin H ml yes 1.46 *Measured at room temperature. VECTASHIELD Hard F Set Mounting Medium measured after hardening. VECTASHIELD is a registered trademark of Vector Laboratories, Inc. 01/15

10 PRODUCT INFORMATION Product VECTASHIELD Hard F Set Mounting Medium with DAPI Catalog No. H-1500 Amount 10 ml Concentration of DAPI Storage Conditions 1.5 µg/ml Refrigerate in the dark VECTASHIELD Hard F Set Mounting Medium preserves fluorescence and hardens after coverslipping. VECTASHIELD Hard F Set Mounting Medium s unique, stable formula prevents rapid photobleaching of fluorescent proteins and fluorescent dyes (see website for compatibility). This special formulation of VECTASHIELD Hard F Set Mounting Medium contains 4,6-diamidino-2-phenylindole (DAPI), which fluoresces when bound to DNA and is used as a nuclear or chromosomal counterstain. This product is designed to be used for in situ hybridization techniques or other procedures requiring fluorescent labeling of DNA. DAPI may also stain RNA but displays greater selectivity for binding DNA over RNA. DAPI excites at about 360 nm and emits at about 460 nm when bound to DNA, producing a blue fluorescence. VECTASHIELD Hard F Set Mounting Medium is supplied in a bottle fitted with a screw cap. A cap fitted with a drop dispenser pipet is also included. The screw cap can be replaced with the drop dispenser cap for routine use and during storage. Once the drop dispenser cap has been secured on the bottle, the bottle should be stored in an upright position. To mount tissues or cells on a slide, dispense VECTASHIELD Hard F Set Mounting Medium onto the specimen. The special pipet supplied with the product is designed to deliver small drop volumes of approximately 25 µl. Coverslip and allow VECTASHIELD Hard F Set Mounting Medium to disperse over the entire specimen. After approximately 15 minutes at room temperature, the coverslip will become immobilized, and optimal antifade ability and refractive index will be achieved. After curing at room temperature for 15 minutes, slides can be placed at 4 ºC, and the mounting media will harden completely overnight. Mounted slides should be stored at 4 C or -20 ºC, protected from light. For prolonged storage, -20 ºC is recommended. If retraction occurs during prolonged storage, remove coverslip and remount. Coverslips can be easily removed by soaking in PBS overnight. (over) Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA U.S.A. Tel: (650) Fax: (650) vector@vectorlabs.com Web site:

11 Notes: When mounting thick sections, adjustments to the mounting protocol may be required. Use of non-hardening VECTASHIELD Mounting Medium (H-1200) may be preferred. The nuclear counterstains, DAPI and PI, will diffuse out of nuclei over time if the cells have been fixed in 100% methanol. In this case non-hardening VECTASHIELD Mounting Medium may be preferred. Note: DAPI is a mutagen and appropriate precautions should be taken when using it. VECTASHIELD Mounting Media Products: Product Counterstain Cat. No. Unit Size Hardening Refractive Index* VECTASHIELD Mounting Medium none H ml no 1.45 DAPI H ml no 1.45 PI H ml no 1.45 VECTASHIELD HardFSet Mounting Medium none H ml yes 1.46 DAPI H ml yes 1.46 TRITC-Phalloidin H ml yes 1.46 *Measured at room temperature. VECTASHIELD Hard F Set Mounting Medium measured after hardening. VECTASHIELD is a registered trademark of Vector Laboratories, Inc. 01/15

12 PRODUCT INFORMATION Product VECTASHIELD Hard F Set Mounting Medium with TRITC-phalloidin Catalog No. H-1600 Amount 10 ml Concentration of TRITC-phalloidin 0.2 µg/ml Storage Conditions Refrigerate in the dark VECTASHIELD Hard F Set Mounting Medium preserves fluorescence and hardens after coverslipping. VECTASHIELD Hard F Set Mounting Medium s unique, stable formula prevents rapid photobleaching of fluorescent proteins and fluorescent dyes (see website for compatibility). This special formulation of VECTASHIELD Hard F Set Mounting Medium contains TRITC-phalloidin. Phalloidin is a bicyclic heptapeptide that specifically binds at the interface between the F-actin subunits. Fluorescent derivatives of phalloidin are used to stain actin filaments. TRITC (tetramethylrhodamine) is excited at 544 nm and emits at 572 nm, producing an orange-red fluorescence. VECTASHIELD Hard F Set Mounting Medium is supplied in a bottle fitted with a screw cap. A cap fitted with a drop dispenser pipet is also included. The screw cap can be replaced with the drop dispenser cap for routine use and during storage. Once the drop dispenser cap has been secured on the bottle, the bottle should be stored in an upright position. To mount tissues or cells on a slide, dispense VECTASHIELD Hard F Set Mounting Medium onto the specimen. The pipet supplied with the product is designed to deliver small drop volumes of approximately 25 µl. Coverslip and allow VECTASHIELD Hard F Set Mounting Medium to disperse over the entire specimen. After approximately 15 minutes at room temperature, the coverslip will become immobilized, and optimal antifade ability and refractive index will be achieved. After curing at room temperature for 15 minutes, slides can be placed at 4 ºC, and the mounting media will harden completely overnight. Mounted slides should be stored at 4 C or -20 ºC, protected from light. For prolonged storage, -20 ºC is recommended. If retraction occurs during prolonged storage, remove coverslip and remount. Coverslips can be easily removed after hardening by soaking in PBS overnight. (over) Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA U.S.A. Tel: (650) Fax: (650) vector@vectorlabs.com Web site:

13 Notes: It is possible to get both cytoskeletal and nuclear counterstaining by mixing this mounting media with VECTASHIELD Hard F Set with DAPI (H-1500) at a 1:1 ratio. DAPI is a nuclear dye that is excited at 360 nm and emits at 460 nm, producing a blue fluorescence. Mix gently to avoid producing bubbles in the mixture and apply to the specimen as described above. When mounting thick sections, adjustments to the mounting protocol may be required. Note: Phalloidin is a toxin and should be handled with care. VECTASHIELD Mounting Media Products: Product Counterstain Cat. No. Unit Size Hardening Refractive Index* VECTASHIELD Mounting Medium none H ml no 1.45 DAPI H ml no 1.45 PI H ml no 1.45 VECTASHIELD HardFSet Mounting Medium none H ml yes 1.46 DAPI H ml yes 1.46 TRITC-Phalloidin H ml yes 1.46 *Measured at room temperature. VECTASHIELD Hard F Set Mounting Medium measured after hardening. VECTASHIELD is a registered trademark of Vector Laboratories, Inc. 01/15

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