Ntb nuclear track emulsion

Manufactured by Kodak

Sourced in United States

The NTB nuclear track emulsion is a lab equipment product designed for the detection and analysis of nuclear particles. It is a photographic emulsion that records the tracks of charged particles, enabling their visualization and study. The core function of the NTB nuclear track emulsion is to provide a medium for the detection and analysis of nuclear interactions and processes.

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Lab products found in correlation

Maxiscript transcription kit, by Thermo Fisher Scientific (3 mentions) Superfrost plus, by Thermo Fisher Scientific (3 mentions) Mrna locator kit, by Thermo Fisher Scientific (2 mentions) Dektol developer, by Kodak (1 mentions) Biomax mr, by Kodak (1 mentions) 35s utp, by PerkinElmer (1 mentions) Taq dna polymerase, by Thermo Fisher Scientific (1 mentions) Bisbenzimide, by Merck Group (1 mentions) Superfrost ultra plus, by Thermo Fisher Scientific (1 mentions) Maxiscript in vitro transcription kit, by Thermo Fisher Scientific (1 mentions)

7 protocols using ntb nuclear track emulsion

In Situ Hybridization of Zebra Finch Brain

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Serial coronal sections (12 μm thick) were cut from 31 whole zebra finch brains using a cryostat, mounted on positively charged slides (SuperfrostPlus^®, Fisher Scientific, Pittsburgh, PA, United States), dried, and stored at −80°C until use. The brain sections were collected in such a way that consecutive sections were mounted on 18 parallel slides. For in situ hybridisation, [³⁵S]UTP-labelled riboprobes were generated from the DNA probes using a MAXIscript transcription kit (Ambion, Austin, TX, United States).
The preparation of tissue was performed using an mRNAlocator Kit (Ambion), according to the manufacturer’s instructions. Tissue was prepared using an mRNA-locator Kit (Ambion) according to manufacturer’s instructions. For hybridisation, we used 80 μl hybridisation buffer and 1 million DPM of labelled probe per slide. Washing procedures included a 30 min incubation in RNase A, followed by decreasing concentrations of sodium-citrate buffer (pH = 7.4) at room temperature, and then at 65°C. After drying, slides were dipped in NTB nuclear track emulsion (Eastman Kodak, Rochester, NY, United States), stored for 3 weeks at 4°C for autoradiography, developed with Kodak Dektol developer, fixed with Kodak fixer, counterstained with Giemsa, and coverslipped with Cytoseal 60 (Stephens Scientific, Riverdale, NJ, United States).

Zachar G., Montagnese C., Fazekas E.A., Kemecsei R.G., Papp S.M., Dóra F., Renner É., Csillag A., Pogány Á, & Dobolyi A. (2020). Brain Distribution and Sexually Dimorphic Expression of Amylin in Different Reproductive Stages of the Zebra Finch (Taeniopygia guttata) Suggest Roles of the Neuropeptide in Song Learning and Social Behaviour. Frontiers in Neuroscience, 13, 1401.

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In Situ Hybridization for Gpr161 Gene Expression

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Both antisense and sense radiolabeled riboprobes for
Gpr161 were synthesized in vitroaccording to the manufacturer’s specifications (Ambion) and labeled
with ³⁵S-UTP (>1,000 Ci/mmol; NEG039H, PerkinElmer LAS
Canada, Inc.). Tissues were cut into 9 μm sections and mounted on
Superfrost^® Plus slides (Fisher Scientific,
12-550-15). Before ISH, sections were fixed in 4% PFA in PBS, treated with
triethanolamine/acetic anhydride, washed and dehydrated in a series of
ethanols. Sections were hybridized overnight at 55°C in 50% deionized
formamide, 0.3 M NaCl, 20mM Tris-HCl pH 7.4, 5 mM EDTA, 10 mM
NaH₂PO₄, 10% dextran sulphate, 1X
Denhardt’s, 50 μg/ml total yeast RNA, and 50-80,000
cpm/μl ³⁵S-labeled cRNA probe. The tissues were subjected
to stringent washing at 65°C in 50% formamide, 2X SSC, 10 mM DTT and
washed in PBS before treatment with 20 μg/ml RNAse A at 37°C
for 30 minutes. Following washes in 2X SSC and 0.1X SSC for 10 minutes at
37°C, the slides were dehydrated and exposed to Kodak BioMaxMR x-ray
film for 4 days then dipped in Kodak NTB nuclear track emulsion and exposed
in light-tight boxes with desiccant at 4°C for 17 days. Photographic
development was carried out in Kodak D-19 [54 (link)]. Sections were counterstained lightly with cresyl violet and
analyzed using brightfield and darkfield microscopy. Sense (control)
riboprobes established the level of background signal.

Hwang S.H., Somatilaka B.N., Badgandi H., Palicharla V.R., Walker R., Shelton J.M., Qian F, & Mukhopadhyay S. (2019). Tulp3 regulates renal cystogenesis by trafficking of cystoproteins to cilia. Current biology : CB, 29(5), 790-802.e5.

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Expression Analysis of Ccdc88b Gene

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The expression of the Ccdc88b gene was monitored by whole mount in situ hybridization. PCR amplification of the designed Ccdc88b probe was performed using primers: 5′-GCGCTATAATACGACTCACTATAGGGAGATCCGAATCTTTGGACCTGCCTTCT-3′ and 3′-GCATTAATTTAGGTGACACTATAGAAGCGAAGCTAGCCGTATCCACTGCTTCA-5′, which contain embedded T7 and SP6 polymerases promoter sites (underlined). ³⁵S-UTP labeled cRNA was synthesized in the presence of SP6 (antisense) and T7 (sense) RNA polymerases, and using ³⁵S-UTP (>1,000 Ci/mmol; PerkinElmer). Tissues were fixed in 4% formaldehyde and hybridized with ³⁵S-labeled cRNA antisense and sense probes. The final slides were dehydrated, and then dipped in Kodak NTB nuclear track emulsion, and exposed for 18 d. The slides were lightly counterstained with cresyl violet and analyzed under both light and dark field optics.
For reverse transcription PCR experiments, total spleen RNA was reverse transcribed into cDNA using oligo-d(T) primers and M-MuLV reverse transcription (Invitrogen). PCR amplification of exons 21–22 of Ccdc88b was performed using primers 5′-CCTGCAGGCTGAAAAGTCA-3′ and 5′-GCTCTCGTCGCTCTCATGGA-3′, and Taq DNA polymerase (Invitrogen), for 28 cycles at 30 s/94°C, 30 s/58°C and 30 s/72°C).

Kennedy J.M., Fodil N., Torre S., Bongfen S.E., Olivier J.F., Leung V., Langlais D., Meunier C., Berghout J., Langat P., Schwartzentruber J., Majewski J., Lathrop M., Vidal S.M, & Gros P. (2014). CCDC88B is a novel regulator of maturation and effector functions of T cells during pathological inflammation. The Journal of Experimental Medicine, 211(13), 2519-2535.

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Fluorescent Visualization of Isotopic Signals

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Following posthybridization, the biotin tyramide deposits were reacted with streptavidin conjugated to the red fluorochrome Cy3 (Jackson; 1:1000; 1 h). Sections were then mounted onto silanized microscope slides, air-dried and processed for autoradiography. To visualize the isotopic signal, the sections were coated with Kodak NTB nuclear track emulsion (Kodak; Rochester, NY, USA; diluted 1:1 with distilled water) which was exposed for 4 weeks, then developed with Kodak processing chemicals. Sections underwent a counterstaining procedure with 10 μg/ml Hoechst (in 0.1 M PBS, pH 7.4, 1 min; Bisbenzimide, Sigma) to enable the distinction of hypothalamic nuclei. After a short rinse in PBS (1 min), the slides were dehydrated in 70, 96, and 100% ethanol (5 min each), cleared with xylene (2 × 5 min), and coverslipped with DPX.

Kalló I., Molnár C.S., Szöke S., Fekete C., Hrabovszky E, & Liposits Z. (2015). Area-specific analysis of the distribution of hypothalamic neurons projecting to the rat ventral tegmental area, with special reference to the GABAergic and glutamatergic efferents. Frontiers in Neuroanatomy, 9, 112.

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Autoradiographic Analysis of DMPFC Brain Tissue

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Two freshly frozen DMPFC brain blocks of subjects were used: one from a 75-year-old female control subject with negative clinical reports for any major diseases and one from a 72-year-old female suicidal individual. Using a cryostat, serial coronal sections (12 μm) were cut and mounted on positively charged slides (SuperfrostPlus^®, Fisher Scientific), dried and stored at −80 °C until use. Further steps were performed according to the procedure described previously by Dobolyi et al. [153 (link)]. Briefly, antisense [35S]UTP-labeled riboprobes were generated from the above-described DNA probes using T7 RNA polymerase of the MAXIscript Transcription Kit (Ambion, Austin, TX, USA) and used for hybridization at 1 million DPM (discharges per minute) activity per slide. Washing procedures included a 30 min incubation in RNase A, followed by decreasing concentrations of sodium-citrate buffer (pH 7.4) at room temperature and subsequently at 65 °C. Following hybridization and washes, slides were dipped in NTB nuclear track emulsion (Eastman Kodak) and stored at 4 °C for 3 weeks for autoradiography. Then, the slides were developed and fixed with Kodak Dektol developer and Kodak fixer, respectively, counterstained with Giemsa and coverslipped with Cytoseal 60 (Stephens Scientific, Kalamazoo, MI, USA).

Dóra F., Renner É., Keller D., Palkovits M, & Dobolyi Á. (2022). Transcriptome Profiling of the Dorsomedial Prefrontal Cortex in Suicide Victims. International Journal of Molecular Sciences, 23(13), 7067.

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In situ Hybridization of C1qbp in Mouse Brain

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To describe the expression pattern of C1qbp in the brain, the fresh tissue was quickly frozen on dry ice. In situ hybridization histochemistry was processed as described previously^{72 (link)}. Briefly, serial coronal sections (12 µm) were cut using a cryostat from bregma level +3.5 mm to −15 mm, mounted on positively charged slides (SuperfrostUltraPlus™; Thermo Fisher Scientific, Pittsburgh, PA, USA), dried, and stored at −80 °C until use. Antisense [35 S]UTP-labelled riboprobes were generated using T7 RNA polymerase of the MAXIscript in vitro transcription kit (Ambion, Austin, TX) from PCR-amplified fragments of the cDNA subcloned into TOPO TA vectors.
Tissue was prepared using an mRNA-locator Kit (Ambion) according to manufacturer’s instructions. For hybridization, we used 80 µl hybridization buffer and 1 million DPM of labelled probe per slide. Washing procedures included a 30 min incubation in RNase A, followed by decreasing concentrations of sodium-citrate buffer (pH = 7.4) at room temperature, and then at 65 °C. After drying, slides were dipped in NTB nuclear track emulsion (Eastman Kodak, Rochester, NY), stored for 3 weeks at 4 °C for autoradiography. Then, the slides were developed and fixed with Kodak Dektol developer and Kodak fixer, respectively, counterstained with Giemsa, dehydrated, and coverslipped with Cytoseal 60 (Stephens Scientific).

Barna J., Dimén D., Puska G., Kovács D., Csikós V., Oláh S., Udvari E.B., Pál G, & Dobolyi Á. (2019). Complement component 1q subcomponent binding protein in the brain of the rat. Scientific Reports, 9, 4597.

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In Situ Hybridization of Hypothalamic Neuropeptides

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Eleven fresh-frozen hypothalamic brain blocks of subjects were used. Using a cryostat, serial coronal sections (12 μm) were cut and mounted on positively-charged slides (SuperfrostPlus^®, Fisher Scientific), dried and stored at −80 °C until use. Further steps were performed according to the procedure described previously [54 (link)]. Briefly, antisense [35S] UTP-labelled riboprobes were generated from the above-described DNA probes using T7 RNA polymerase of the MAXIscript Transcription Kit (Ambion, Austin, TX, USA) and used for hybridization at 1 million DPM (discharges per minute) activity per slide. Washing procedures included a 30 min incubation in RNase A followed by decreasing concentrations of sodium-citrate buffer (pH 7.4) at room temperature and subsequently at 65 °C. Following hybridization and washes, slides were dipped in NTB nuclear track emulsion (Eastman Kodak) and stored at 4 °C for 3 weeks (GLP-1R), or 1 day (Oxy) for autoradiography. Then, the slides were developed and fixed with Kodak Dektol developer and Kodak fixer, respectively, counterstained with Giemsa and coverslipped with Cytoseal 60 (Stephens Scientific, Kalamazoo, MI, USA). The density of autoradiographic signal was calculated with Image J software following microscopic photography in the dark-field using 40X objective.

Renner É., Dóra F., Oszwald E., Dobolyi Á, & Palkovits M. (2022). Elevated Glucagon-like Peptide-1 Receptor Level in the Paraventricular Hypothalamic Nucleus of Type 2 Diabetes Mellitus Patients. International Journal of Molecular Sciences, 23(24), 15945.

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