in vitro

Examples of in vitro in the following topics:

• In Vivo Testing

  • In vivo testing follows strict guidelines and humane animal use ethics.
  • Polyclonal antibodies have various applications in the clinic and in research laboratories.
  • In addition to having bodies that work similar to humans and other animals, rodents are small in size, easy to handle, relatively inexpensive to buy and keep, and produce many offspring in a short period of time.
  • In vivo testing remains a crucial step for the evaluation of in vitro experimental findings and the production of immunological solutions needed for the diagnosis of human diseases.
  • Animals are used in laboratory experiments to translate in vitro findings.

• General Staining Methods

  • Staining is a technique used in microscopy to enhance contrast in a microscopic image.
  • In-vivo staining is the process of dyeing living tissue -- in vivo means "in life" (as contrasted to in-vitro staining).
  • In-vitro staining involves coloring cells or structures that have been removed from their biological context.
  • What follows here are some common aspects of the process of preparing for in-vitro staining.
  • In some cases, cells may be grown directly on a slide.

• Ribosomes

  • Proteins synthesized in each of these locations serve a different role in the cell.
  • In prokaryotes, ribosomes can be found in the cytosol as well.
  • In most bacteria, the most numerous intracellular structure is the ribosome which is the site of protein synthesis in all living organisms.
  • However, the reconstitution intermediates are not the same as in vitro.
  • The intermediates in the in vivo assembly are precursor rRNA which is different from in vitro which uses matured rRNA.

• Microarrays and the Transciptome

  • Analysis of the transcriptomes of human oocytes and embryos is used to understand the molecular mechanisms and signaling pathways controlling early embryonic development, and could theoretically be a powerful tool in making proper embryo selection during in vitro fertilization.
  • The term can be applied to the total set of transcripts in a given organism, or to the specific subset of transcripts present in a particular cell type.
  • A number of organism-specific transcriptome databases have been constructed and annotated to aid in the identification of genes that are differentially expressed in distinct cell populations.
  • DNA microarrays can provide a genome-wide method for comparison of the abundance of DNAs in the same samples.The DNA in spots can only be PCR products specific for individual genes.
  • Analysis of the transcriptomes of human oocytes and embryos is used to understand the molecular mechanisms and signaling pathways controlling early embryonic development, and could theoretically be a powerful tool in making proper embryo selection during in vitro fertilization.

• DNA Sequencing Techniques

  • By using a predetermined ratio of deoxyribonucleotides to dideoxynucleotides, it is possible to generate DNA fragments of different sizes when replicating DNA in vitro.
  • A Sanger sequencing reaction is just a modified in vitro DNA replication reaction.
  • Most of the time in a Sanger sequencing reaction, DNA Polymerase will add a proper dNTP to the growing strand it is synthesizing in vitro.
  • If enough template DNAs are included in the reaction mix, each one will have the ddNTP inserted at a different random location, and there will be at least one DNA terminated at each different nucleotide along its length for as long as the in vitro reaction can take place (about 900 nucleotides under optimal conditions.)
  • In shotgun sequencing, multiple copies of the same chromosome are isolated and then fragmented in random locations.

• Fertilization

  • In medicine, this process is referred to as fertilization; in lay terms, it is more commonly known as "conception."
  • However, the advent of artificial insemination and in vitro fertilization have made achieving pregnancy possible without engaging in sexual intercourse.
  • The process of fertilization occurs in several steps; the interruption of any of them can lead to failure.
  • The sperm must undergo capacitation in the female's reproductive tract over several hours, which increases its motility and destabilizes its membrane.
  • The sperm and the egg cell (which has been released from one of the female's two ovaries) unite in one of the two fallopian tubes.

• Primer Extension Analysis

  • Second, it can be used to quantify the amount of transcript in an in vitro transcription system.
  • Third, it can be used to determine the locations of breaks or modified bases in a mixed population of RNA or DNA samples.
  • This is useful in applications like footprinting.
  • In one, the modified nucleotide cannot be recognized by the polymerase or reverse transcriptase; in such cases, the chain ends at the site of modification.
  • In the other, the modification is converted in a later step of the analysis to a strand break by chemical treatment.

• Infertility

  • These include diseases (such as sexually-transmitted diseases that can cause scarring of the reproductive tubes in either men or women) or developmental problems frequently related to abnormal hormone levels in one of the individuals.
  • A common type of assisted reproductive technology is in vitro fertilization (IVF) where an egg and sperm are combined outside the body and then placed in the uterus.
  • IVF procedures produce a surplus of fertilized eggs and embryos that can be frozen and stored for future use; the procedures can also result in multiple births.
  • Define infertility and discuss ways in which it can be treated

• Cultivation of Specimen

  • It is also critical that the pathogen is isolated in a pure culture first.
  • In most cases, specimens are also inoculated into differential media that define such characteristics as fermentation patters (mannitol salt and MacConkey agar) and as reactions in blood (blood agar).
  • Serological testing uses in-vitro diagnostic testing of serum, has a high degree of specificity and sensitivity, and is based on the specificity an antibody has for its antigen.
  • Results of specimen analysis are entered in the patient's summary chart.
  • This blood is cultured in a bottle to detect bloodstream infections.

• Recombinant DNA Technology

  • Molecular cloning permits the replication of a specific DNA sequence in a living microorganism.
  • The fundamental difference between the two methods is that molecular cloning involves replication of the DNA in a living microorganism, while PCR replicates DNA in an in vitro solution, free of living cells.
  • The creation of recombinant DNA is in many ways the simplest step of the molecular cloning process.
  • The DNA mixture, previously manipulated in vitro, is moved back into a living cell, referred to as the host organism.
  • The competent cells are grown in the presence of X-gal.