Vectashield h 1400

Manufactured by Vector Laboratories

Sourced in United States

Vectashield H-1400 is a water-based mounting medium designed for fluorescence microscopy. It is formulated to preserve fluorescent signals and reduce photobleaching of fluorescent dyes.

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Lab products found in correlation

Apotome, by Zeiss (6 mentions) Vt1200, by Leica (2 mentions) Paraformaldehyde pfa, by Merck Group (2 mentions) Permount, by Thermo Fisher Scientific (1 mentions) Vectastain elite abc kit, by Vector Laboratories (1 mentions) Photo flo, by Kodak (1 mentions) Alexa fluor 488 conjugate, by Thermo Fisher Scientific (1 mentions) Fv10i, by Olympus (1 mentions) Alexa fluor 488 secondary antibody, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 donkey anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 donkey anti mouse igg h l, by Thermo Fisher Scientific (1 mentions) Monoclonal anti gapdh antibody, by Merck Group (1 mentions) Lsm 780 confocal microscope, by Zeiss (1 mentions) Mab386, by Merck Group (1 mentions) Ab9610, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Axioimager z2 fluorescence microscope, by Zeiss (1 mentions) Technovit 8100, by Kulzer (1 mentions)

Close spelling variants of this product

Vectashield (3789 citations) H-1400 (25 citations) Vectashield medium (H-1400; (5 citations) Vectashield hard set mounting medium (H-1400; (5 citations) Vectashield mounting medium (H-1400; (4 citations) VECTASHIELD Hardset Antifade Mounting Medium (H-1400; (3 citations) Vectashield HardSet H-1400 (2 citations)

9 protocols using vectashield h 1400

Perfusion, Fixation, and Sectioning for Fluorescence and Electron Microscopy

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Animals were deeply anesthetized via a lethal dose of sodium pentobarbitone (100 mg/kg, intraperitoneally), and perfused transcardially with 5 ml 1% sodium nitrite prewash in 0.1M phosphate buffered saline followed immediately by 60 ml 4% paraformaldehyde/0.1% glutaraldehyde in 0.1M phosphate buffer. Heads were postfixed overnight after which the brain was dissected from the skull, embedded in gelatin-albumin, and cut in the transverse plane at 50–60 μm using a vibrating microtome (VT1200S; Leica Systems, Nussloch, Germany). All reagents and rinses were made up in 0.12M Tris-buffered saline.
Visualization of Alexa Fluor 488 or 555 did not require any further tissue processing. Coronal sections were mounted and coverslipped using Vectashield (H-1400; Vector Labs) and viewed with a fluorescent microscope. To visualize BDA labeling for brightfield and electron microscopy, sections were incubated for 10 min in 1% H2O2, rinsed 3×, permeabilized for 1 h in 0.5% Photo-Flo (Kodak, Rochester, NY, USA) or 0.5% Triton X-100 (brightfield only), and then incubated in ABC (Vectastain Elite ABC Kit, PK-6100; Vector Labs) with 0.5% Photo-Flo/Triton for 1 h. The tissue was rinsed 3× again and BDA labeling was developed using nickel-intensified diaminobenzidine (DAB; Sigma-Aldrich). Sections were mounted, dehydrated, and coverslipped with Permount (Fisher Scientific, Pittsburgh, PA, USA).

Ryugo D.K, & Milinkeviciute G. (2023). Differential projections from the cochlear nucleus to the inferior colliculus in the mouse. Frontiers in Neural Circuits, 17, 1229746.

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Double Immunostaining of CK18 and Ki-67 in Frozen Tissue

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Double immunofluorescence staining was performed on frozen tissue sections. Antigen
retrieval was carried out using Histo VT One at 70°C for 20 min. The slides were then
cooled to room temperature. Sections were incubated in blocking buffer (5% NHS/1% bovine
serum albumin /0.2% Triton X-100 in PBS) for 30 min at room temperature. The sections were
incubated with anti-CK18 antibody (diluted 1:1,000) overnight at 4°C. After rinsing 3 × 5
min in PBS, the sections were incubated in anti-IgG (H+L) mouse, horse-poly biotin (Vector
Laboratories) diluted at 1:200 for 1 hr. The sections were then incubated in streptavidin,
Alexa Fluor^® 488 conjugate (Invitrogen, Carlsbad, CA, U.S.A.) diluted at 1:200
for 1 hr, followed by three PBS washes. The sections were then incubated with Alexa
Fluor®555 mouse anti-Ki-67 antibody (B56; BD Pharmingen, San Diego, CA, U.S.A.) overnight
at 4°C. The slides were mounted in Antifade medium (Vectashield H-1400, Vector
Laboratories). The stained sections were examined using a confocal microscope (Fv10i;
Olympus, Tokyo, Japan).

HARA A., ABE T., HIRAO A., SANBE K., AYAKAWA H., SARANTONGLAGA B., YAMAGUCHI M., SATO A., KHURCHABILIG A., OGATA K., FUKUMORI R., SUGITA S, & NAGAO Y. (2017). Histochemical properties of bovine and ovine mammary glands during fetal development. The Journal of Veterinary Medical Science, 80(2), 263-271.

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Immunohistochemical Detection of Hippocampal p-p38 Activation

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All mice were deeply anesthetized with ketamine/xylazine and transcardially perfused with 0.1 M phosphate buffer (PB) followed by 4% paraformaldehyde (4% PFA) dissolved in 0.1M PB. Brains were extracted and post-fixed in 4% PFA for 24 h. Brains were transferred to 30% sucrose for 48–72 h before slicing 30 μm coronal sections through the extent of the hippocampus using a cryostat. Sections were stored in cryoprotectant at −20°C until use. For post-processing of hippocampal slices used in electrophysiology experiments, slices were bath fixed in 4% PFA overnight at 4°C prior to incubation in 30% sucrose. 18 μm coronal sections through the extent of the hippocampus were cut and directly mounted onto charged slides and processed for staining. For staining, slices were washed extensively in PBS + 0.1% Triton-X (PBST) before blocking in 5% normal goat serum (NGS) + PBST for 1 hour to reduce non-specific background. Slices were incubated with p-p38 antibodies (9211S, Cell Signaling) overnight at room temperature in 5% NGS + PBST. The next day, slices were rinsed extensively in PBST and incubated in AlexaFluor 488 secondary antibodies (A11008, Invitrogen) for 1.5 h at room temperature. The slices were washed with PBST and counterstained with DAPI to observe nuclei before mounting on slides with Vectashield (H-1400, Vector Labs) to prevent fading.

Darcy M.J., Jin S.X, & Feig L.A. (2014). R-Ras Contributes to LTP and Contextual Discrimination. Neuroscience, 277, 334-342.

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Immunostaining of Genetically Labeled Cells

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Sections from nestin-CreER^T2 × GluCl lines, were immunostained for YFP to enhance the fluorescent signal. Sections from nestinCreER^T2 × Ai3 lines were immunostained for parvalbumin to identify cerebellar and cortical interneurons, calbindin to identify striatal medium spiny neurons and cortical interneurons, S100B to identify astrocytes, and collagen IV to identify vascular endothelial cells. Sections were blocked for 1 hour at room temperature (5% goat serum, 1% BSA, 0.5% Tween- 20 and 0.1% Triton-X 100 in TBS) and then incubated overnight at 4° C with primary antibody diluted in blocking solution. The following day, sections were rinsed with TBS and incubated with Alexa Fluor 488-conjugated (YFP only) or Alexa Fluor 568-conjugated secondary antibody diluted 1:500 in blocking solution for 2 hours at room temperature (Invitrogen, anti-chicken A11039, anti-goat A11057, anti-rabbit A11011). Sections were mounted with aqueous anti-fade mounting medium (Vectashield H-1400, Vector Laboratories) and processed for fluorescent detection.

Sun M.Y., Yetman M.J., Lee T.C., Chen Y, & Jankowsky J.L. (2014). Specificity and efficiency of reporter expression in adult neural progenitors varies substantially among nestin-CreERT2 lines. The Journal of comparative neurology, 522(5), 1191-1208.

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Histological Validation of Neuronal Recording Sites

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Post-hoc histology was performed to confirm the location of the silicon probe. Both silicon probe shanks were coated with DiI (#C7000, Thermo Fisher Scientific). After the recordings, mice were deeply anesthetized with isoflurane then cardiac perfused with phosphate-buffered saline (PBS) followed by 4% Paraformaldehyde (PFA; Sigma-Aldrich). Whole brains were carefully dissected and post-fixed into 4% PFA overnight and sliced using a vibratome (Leica VT1200). Coronal serial sections were made at a thickness of 70μm in a 4 °C PBS solution, mounted on a slide glass, and cover slipped with VECTASHIELD (H-1400, Vector Laboratories). We used the Paxinos Mouse Brain Atlas (Franklin & Paxinos, 2008 ) for nomenclature of brain regions. Images were taken using a widefield microscope (Apotome, Ziess). Contrast, brightness, and pseudocolor were adjusted in Image J (Schneider et al., 2012 (link)). Images were tilted to align to illustration based on the atlas.

Hur S.W., Safaryan K., Yang L., Blair H.T., Masmanidis S.C., Mathews P.J., Aharoni D, & Golshani P. (2023). Correlated signatures of social behavior in cerebellum and anterior cingulate cortex. bioRxiv.

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Immunofluorescence Staining of Mouse Erythrocytes

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Mouse erythrocytes were fixed with 100% ice-cold methanol for 10 min. The fixed cells were washed two times with PBS, blocked by 1% BSA in PBS (Blocking buffer, pH 7.4.) for 1 h at room temperature. Cells were incubated with monoclonal anti-GAPDH antibody (Sigma-Aldrich, 1:100 in blocking buffer) at 4 °C for overnight. The cells were washed three times with PBS, incubated with Alexa fluor 594 donkey anti-mouse IgG(H+L) (1:1,000 dilution) or Alexa fluor 488 donkey anti-rabbit IgG(H+L) (1:1,000 dilution) (Life Technologies) for 1 h at room temperature in dark, then washed three times and re-suspended in PBS. Cell-smear were made and dried in dark. The slides were mounted with cover glass by mounting medium (VECTASHIELD H-1400, Vector, CA). Pictures were taken under Zeiss LSM 780 confocal microscope (Carl Zeiss Inc., Jena, Germany).

Sun K., Zhang Y., D'Alessandro A., Nemkov T., Song A., Wu H., Liu H., Adebiyi M., Huang A., Wen Y.E., Bogdanov M.V., Vila A., O'Brien J., Kellems R.E., Dowhan W., Subudhi A.W., Jameson-Van Houten S., Julian C.G., Lovering A.T., Safo M., Hansen K.C., Roach R.C, & Xia Y. (2016). Sphingosine-1-phosphate promotes erythrocyte glycolysis and oxygen release for adaptation to high-altitude hypoxia. Nature Communications, 7, 12086.

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Immunocytochemistry Protocols for Oligodendrocyte Markers

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For immunocytochemistry, the cells were fixed with 4% paraformaldehyde (PFA)/phosphate-buffered saline (PBS) for 15 min at room temperature; washed with PBS; and blocked for 45 min at room temperature with blocking solution (PBS containing 5% normal donkey serum, 0.5% Triton X-100 and 0.05% sodium azide). The samples were incubated overnight at 4°C with primary antibodies diluted in blocking solution, washed three times in PBS, incubated for 45 min with secondary antibodies and washed in PBS and mounted with mounting medium (Vectashield H-1400, vectorlabs). For primary antibodies, the following antibodies were used: rat monoclonal antibody to MBP (1 : 300, MAB386, Chemicon), mouse hybridoma supernatants to O4 (1 : 5), rabbit polyclonal antibodies to Olig2 (1 : 500, AB9610, Chemicon) and Caspr (1 : 1000) [22 (link)]. Secondary antibodies were obtained from Jackson Immunoresearch. DAPI was obtained from Invitrogen.

Kim J.Y., Yoon J.Y., Sugiura Y., Lee S.K., Park J.D., Song G.J, & Yang H.J. (2019). Dendropanax morbiferus leaf extract facilitates oligodendrocyte development. Royal Society Open Science, 6(6), 190266.

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Visualizing Symbiont Colonization in Parasitoids

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Adult females and larvae of W + R + S. endius were fixed in 4% formaldehyde (in 1× PBS) for 24 h and then embedded in Technovit 8100 (Heraeus Kulzer, Wehrheim, Germany). Semithin sections (8 μm) were obtained on a rotary microtome (Microm HM355S) with a glass blade and transferred to silanized microscope slides. Samples were hybridized for 90 min at 50 °C in hybridization buffer (0.9 M NaCl, 0.02 M Tris/HCl pH 8.0, 0.01% SDS) containing 25 nM of each of the symbiont-specific probes as well as 5 μg/ml DAPI for counterstaining of host cell nuclei. Residual probes were removed by a 20-min wash step at 50 °C with pre-warmed wash buffer (0.1 M NaCl, 0.02 M Tris/HCl pH 8.0, 0.01% SDS, 5 mM EDTA), followed by a 20-min washing step in dH 2 O. After short rinsing in dH 2 O and shaking off the excess liquid, slides were covered with VectaShield H-1400 (Vector, Burlingame, USA) and inspected on an AxioImager.Z2 fluorescence microscope with Apotome (Zeiss, Jena, Germany).
The Rickettsia probe was 5′-Cy5-CCGGCATTACCCGC TGGCAA-3′ [38] (link), and the Wolbachia probe was 5′-Cy3-CTTCTGTGAGTACCGTCATTATC-3′ [39] , both targeting the 16S rRNA. Aposymbiotic parasitoids were used as negative control.

Semiatizki A., Weiss B., Bagim S., Rohkin-Shalom S., Kaltenpoth M, & Chiel E. (2020). Effects, interactions, and localization of Rickettsia and Wolbachia in the house fly parasitoid, Spalangia endius. Microbial ecology, 80(3).

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Histological Validation of Neuronal Recordings

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Post-hoc histology was performed to confirm the location of the silicon probe. Both silicon probe shanks were coated with DiI (#C7000, Thermo Fisher Scientific). After the recordings, mice were deeply anesthetized with isoflurane then cardiac perfused with phosphate-buffered saline (PBS) followed by 4% Paraformaldehyde (PFA; Sigma-Aldrich). Whole brains were carefully dissected and post-fixed into 4% PFA overnight and sliced using a vibratome (Leica VT1200). Coronal serial sections were made at a thickness of 70 μm in a 4 °C PBS solution, mounted on a slide glass, and cover slipped with VECTASHIELD (H-1400, Vector Laboratories). We used the Paxinos Mouse Brain Atlas (Franklin and Paxinos, 2008 ) for nomenclature of brain regions. Images were taken using a widefield microscope (Apotome, Ziess). Contrast, brightness, and pseudocolor were adjusted in Image J (Schneider et al., 2012 (link)). Images were tilted to align to illustration based on the atlas.

Hur S.W., Safaryan K., Yang L., Blair H.T., Masmanidis S.C., Mathews P.J., Aharoni D, & Golshani P. (2024). Correlated signatures of social behavior in cerebellum and anterior cingulate cortex. eLife, 12, RP88439.

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