1. The document provides a 13-step immunohistochemistry protocol for slide-mounted paraffin sections. It details the steps for deparaffinizing, antigen retrieval, inactivation of endogenous peroxidase, primary and secondary antibody incubation, signal detection using DAB, hematoxylin counterstaining, dehydration, mounting, and imaging of samples.
2. Key steps include deparaffinizing tissue sections using xylene and graded alcohols, optional antigen retrieval by heating sections in citric acid buffer, blocking endogenous peroxidase with hydrogen peroxide, incubating with primary and secondary antibodies, developing signal using DAB substrate, counterstaining with hematoxylin, and dehydrating
This presentation includes the basic knowledge of Karyotyping with a lot of understandable knowledge and also how to use it properly. I hope all the finders liked it and also remember me in your precious Dua. Thank You!
It is generally recognized that stained fecal films are the single most productive means of stool examination for intestinal protozoa. The permanent stained smear facilitates detection and identification of cysts and trophozoites and affords a permanent record of the protozoa encountered. Small protozoa, missed by wet mount examinations (of either unconcentrated or concentrated samples) are often seen on the stained smear. The Wheatley Trichrome technique for fecal specimens is a modification of Gomori's original staining procedure for tissue. It is a rapid, simple procedure, which produces uniformly well-stained smears of the intestinal protozoa, human cells, yeast, and artifact material.
This presentation includes the basic knowledge of Karyotyping with a lot of understandable knowledge and also how to use it properly. I hope all the finders liked it and also remember me in your precious Dua. Thank You!
It is generally recognized that stained fecal films are the single most productive means of stool examination for intestinal protozoa. The permanent stained smear facilitates detection and identification of cysts and trophozoites and affords a permanent record of the protozoa encountered. Small protozoa, missed by wet mount examinations (of either unconcentrated or concentrated samples) are often seen on the stained smear. The Wheatley Trichrome technique for fecal specimens is a modification of Gomori's original staining procedure for tissue. It is a rapid, simple procedure, which produces uniformly well-stained smears of the intestinal protozoa, human cells, yeast, and artifact material.
Immunohistochemistry Antibody Validation Report for Anti-Phospho-JNK1/2/3 (T1...St John's Laboratory Ltd
Serine/threonine-protein kinase involved in various processes such as cell proliferation, differentiation, migration, transformation and programmed cell death. Extracellular stimuli such as proinflammatory cytokines or physical stress stimulate the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. In this cascade, two dual specificity kinases MAP2K4/MKK4 and MAP2K7/MKK7 phosphorylate and activate MAPK8/JNK1. In turn, MAPK8/JNK1 phosphorylates a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. Phosphorylates the replication licensing factor CDT1, inhibiting the interaction between CDT1 and the histone H4 acetylase HBO1 to replication origins. Loss of this interaction abrogates the acetylation required for replication initiation.
Anti-Phospho-JNK1/2/3 (T183/Y185)-http://www.stjohnslabs.com/phospho-jnk123-t183y185-antibody
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Pharmaceutical Quality Management of Dexamethasone tablets BP
Dexamethasone tablets USP
DEXAMETHSONE OPTHALMIC SUSPENSION BP
DEXAMETHSONE OPTHALMIC SUSPENSION USP
Dexamethasone is a synthetic (man-made) corticosteroid.
Corticosteroids are naturally-occurring chemicals produced by the adrenal glands located above the kidneys.
Aquaporin 4 also known as AQP4 is a protein which in humans is encoded by the AQP4 gene. AQP4 belongs to the aquaporin family of integral membrane proteins that conduct water through the cell membrane.
AQP4 is constitutively expressed in the basolateral cell membrane of principal collecting duct cells in the kidney and provide a pathway for water to exit these cells.
AQP4 is also expressed in astrocytes and is upregulated by direct insult to the central nervous system.
Anti-Aquaporin 4 -http://www.stjohnslabs.com/aquaporin-4-antibody-p-98599
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Immunohistochemistry Antibody Validation Report for Anti-Phospho-JNK1/2/3 (T1...St John's Laboratory Ltd
Serine/threonine-protein kinase involved in various processes such as cell proliferation, differentiation, migration, transformation and programmed cell death. Extracellular stimuli such as proinflammatory cytokines or physical stress stimulate the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. In this cascade, two dual specificity kinases MAP2K4/MKK4 and MAP2K7/MKK7 phosphorylate and activate MAPK8/JNK1. In turn, MAPK8/JNK1 phosphorylates a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. Phosphorylates the replication licensing factor CDT1, inhibiting the interaction between CDT1 and the histone H4 acetylase HBO1 to replication origins. Loss of this interaction abrogates the acetylation required for replication initiation.
Anti-Phospho-JNK1/2/3 (T183/Y185)-http://www.stjohnslabs.com/phospho-jnk123-t183y185-antibody
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Pharmaceutical Quality Management of Dexamethasone tablets BP
Dexamethasone tablets USP
DEXAMETHSONE OPTHALMIC SUSPENSION BP
DEXAMETHSONE OPTHALMIC SUSPENSION USP
Dexamethasone is a synthetic (man-made) corticosteroid.
Corticosteroids are naturally-occurring chemicals produced by the adrenal glands located above the kidneys.
Aquaporin 4 also known as AQP4 is a protein which in humans is encoded by the AQP4 gene. AQP4 belongs to the aquaporin family of integral membrane proteins that conduct water through the cell membrane.
AQP4 is constitutively expressed in the basolateral cell membrane of principal collecting duct cells in the kidney and provide a pathway for water to exit these cells.
AQP4 is also expressed in astrocytes and is upregulated by direct insult to the central nervous system.
Anti-Aquaporin 4 -http://www.stjohnslabs.com/aquaporin-4-antibody-p-98599
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Sterilisation-
Is the process of making something free from bacteria or other living microorganisms.
Sterility Testing-
Are done to detect if viable forms of micro-organisms are present or not on or in the pharmaceutical preparations.
Transcription factor that is the main target of insulin signaling and regulates metabolic homeostasis in response to oxidative stress. Binds to the insulin response element (IRE) with consensus sequence 5'-TT[G/A]TTTTG-3' and the related Daf-16 family binding element (DBE) with consensus sequence 5'-TT[G/A]TTTAC-3'. Activity suppressed by insulin. Main regulator of redox balance and osteoblast numbers and controls bone mass. Orchestrates the endocrine function of the skeleton in regulating glucose metabolism. Acts synergistically with ATF4 to suppress osteocalcin/BGLAP activity, increasing glucose levels and triggering glucose intolerance and insulin insensitivity. Also suppresses the transcriptional activity of RUNX2, an upstream activator of osteocalcin/BGLAP. In hepatocytes, promotes gluconeogenesis by acting together with PPARGC1A and CEBPA to activate the expression of genes such as IGFBP1, G6PC and PCK1. Important regulator of cell death acting downstream of CDK1, PKB/AKT1 and SKT4/MST1. Promotes neural cell death. Mediates insulin action on adipose tissue. Regulates the expression of adipogenic genes such as PPARG during preadipocyte differentiation and, adipocyte size and adipose tissue-specific gene expression in response to excessive calorie intake. Regulates the transcriptional activity of GADD45A and repair of nitric oxide-damaged DNA in beta-cells. Required for the autophagic cell death induction in response to starvation or oxidative stress in a transcription-independent
Anti-FoxO1-http://www.stjohnslabs.com/foxo1-antibody-p-92348
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Promotes microtubule assembly and stability, and might be involved in the establishment and maintenance of neuronal polarity. The C-terminus binds axonal microtubules while the N-terminus binds neural plasma membrane components, suggesting that tau functions as a linker protein between both. Axonal polarity is predetermined by TAU/MAPT localization (in the neuronal cell) in the domain of the cell body defined by the centrosome. The short isoforms allow plasticity of the cytoskeleton whereas the longer isoforms may preferentially play a role in its stabilization.
Anti-Tau- http://www.stjohnslabs.com/anti-tau-antibody?filter_name=STJ98827
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Involved in redox regulation of the cell. Reduces peroxides with reducing equivalents provided through the thioredoxin system but not from glutaredoxin. May play an important role in eliminating peroxides generated during metabolism. Might participate in the signaling cascades of growth factors and tumor necrosis factor-alpha by regulating the intracellular concentrations of H2O2. Reduces an intramolecular disulfide bond in GDPD5 that gates the ability to GDPD5 to drive postmitotic motor neuron differentiation (By similarity). / 2 R'-SH + ROOH = R'-S-S-R' + H2O + ROH.
Anti-Peroxiredoxin 1-http://www.stjohnslabs.com/peroxiredoxin-1-antibody-p-98649
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Promotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2 (By similarity). Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.
Anti-Bad-http://www.stjohnslabs.com/bad-antibody-p-91313
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Type II collagen is specific for cartilaginous tissues. It is essential for the normal embryonic development of the skeleton, for linear growth and for the ability of cartilage to resist compressive forces.
Anti-COL2A1 -http://www.stjohnslabs.com/col2a1-antibody-p-91775
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation.
Anti-Histone H2A.X-http://www.stjohnslabs.com/histone-h2ax-antibody-p-92631
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Anti-Histone H3-http://www.stjohnslabs.com/histone-h3-antibody-p-92641
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Since no universal tissue preparation method will be ideal for all sample and tissue types, the IHC protocol given here is intended as a starting point from which the experimenter should optimize as needed.
This guide is key to successful IHC experiments. Since no universal tissue preparation method will be ideal for all sample and tissue types, all protocols given here are intended as a starting point from which the experimenter must optimize as needed.
Immunohistochemistry Antibody Validation Report for Anti-Phospho-Stat1 (Y701)...St John's Laboratory Ltd
Signal transducer and transcription activator that mediates cellular responses to interferons (IFNs), cytokine KITLG/SCF and other cytokines and other growth factors. Following type I IFN (IFN-alpha and IFN-beta) binding to cell surface receptors, signaling via protein kinases leads to activation of Jak kinases (TYK2 and JAK1) and to tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize and associate with ISGF3G/IRF-9 to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of IFN-stimulated genes (ISG), which drive the cell in an antiviral state. In response to type II IFN (IFN-gamma), STAT1 is tyrosine- and serine-phosphorylated. It then forms a homodimer termed IFN-gamma-activated factor (GAF), migrates into the nucleus and binds to the IFN gamma activated sequence (GAS) to drive the expression of the target genes, inducing a cellular antiviral state.
Anti-Phospho-Stat1 (Y701)-http://www.stjohnslabs.com/phospho-stat1-y701-antibody
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection.
Anti-Cytokeratin 18-http://www.stjohnslabs.com/cytokeratin-18-antibody-p-91959
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Transthyretin (TTR) is a transport protein in the serum and cerebrospinal fluid that carries the thyroid hormone thyroxine (T4) and retinol-binding protein bound to retinol. This is how transthyretin gained its name: transports thyroxine and retinol. The liver secretes transthyretin into the blood, and the choroid plexus secretes TTR into the cerebrospinal fluid.
TTR was originally called prealbumin(or thyroxine-binding prealbumin) because it ran faster than albumin on electrophoresis gels.
Anti-TTR - http://www.stjohnslabs.com/anti-ttr-antibody?filter_name=STJ98876
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Similar to Immunohistochemistry Guide for Slide Mounted Paraffin Sections (20)
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Immunohistochemistry Guide for Slide Mounted Paraffin Sections
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1
Immunohistochemistry Guide for Slide-mounted Paraffin Sections
Deparaffinizing and rehydration
1. Immerse the slides in Xylene Ⅰ and Xylene Ⅱ successively for 10 minutes respectively.
2. Immerse the slides in anhydrous ethanol Ⅰ , anhydrous ethanol Ⅱ , 95% ethanol, 80% ethanol, and 70% ethanol successively for 5
minutes respectively, and then use deionized water to wash the slides for 2 minutes and repeat 2 times.
Antigen retrieval (optional)
3. Put the slides into the repair box, and then add 0.01M citric acid buffer (PH6.0) to make the tissue immersed.
4. Repair the antigen with medium power microwave for 10 minutes (start timing when the liquid boils), and avoid the tissue from drying during
the process.
5. Take out the repair box from microwave oven, and it cool down naturally. Take out the slides when the repair solution cooled down to room
temperature, and wash the slides with PBS (PH7.4) for 3 minutes and repeat 3 times. (Don’t flush the tissue directly during the washing
process in order to avoid breaking up the tissue).
Inactivation
6. Add the prepared 3% H2O2 to the slides drop by drop to block endogenous peroxidase. Then incubate them at room temperature for 15
minutes (dilute 30% H2O2 with methanol or distilled water), finally use PBS to wash the slides with PBS for 3 minutes and repeat 3 times.
Antibody incubation
7. Blot up absorbent paper with PBS, and add 5% normal serum (Sharing the same or similar species with secondary antibodies) drop by drop
on the sections, then block it at 37°C for 30 minutes.
8. Wipe dry the liquid around the tissue on the slides with absorbent paper, and use an oil pen to draw a circle around the tissue, and
then add the diluted primary antibodies drop by drop. Add PBS to the section of controls if the negative controls are required. After adding
primary antibodies, put the slides into wet box to be incubated at 4°C overnight. (The optimum dilution ratio of the antibodies should be
pre-determined through experiments in advance).
9. Wash the slides with PBS for 3 times, each time for 2 minutes, and then add HRP-conjugated secondary antibodies after wiping dry the
slides with absorbent paper, finally incubate the slides at 37°C for 30 minutes.
Signal detection
10. Wash the sections with PBS for 3 minutes and repeat 4 times, and wipe dry the sections with absorbent paper, then add DAB substrate
reagent that is prepared freshly drop by drop to each section, and observe them under a microscope. The positive signal appears brown-
yellow or brown in color. The time should be well controlled to avoid the color appears too deep. Wash the section with tap water to
terminate the reaction.
11. Hematoxylin counterstaining: Immerse slides in Harris hematoxylin solution for about 30 seconds to 1 minute, and then transfer slides into
ethanol solution with 1% HCl after washing with water, finally wash them with water. (Optional)
Dehydration and mounting
12. Firstly immerse the slides in water and wash them, and then put the slides into the following reagents in order to dehydrate and permeate:
70% ethanol, 80% ethanol, 90% ethanol, 95% ethanol, anhydrous ethanol Ⅰ , anhydrous ethanol, Xylene Ⅰ and Xylene Ⅱ . Put the
slides in each reagent for 2 minutes, and finally air dry the sections in the fume cupboard.
13. Drop resinene beside the section, and then cover them with the cover glass. In order to avoid air bubble, firstly lay one side of the cover
glass flat and then gently lay another side flat. Finally dry the sealed sections by laying them in the fume cupboard
14. Observe the dried sections and collect images with a microscope.