SIMPLE STAINING | Bacterial Staining Technique | Microbiology | Vivek Srinivas | #Bacteriology

00:10:55
https://www.youtube.com/watch?v=JHLsb97_wTA

Resumen

TLDRVideoen demonstrerer en enkel bakteriefargingsteknikk, som fokuserer på forberedelse av bakteriesmear og bruk av metylblå eller fortynnet karbolfusjon for visualisering. Den dekker bakterieformer, arrangementer og fargeprosessen, inkludert varmefiksering og mikroskopisk undersøkelse. Hovedformene for bakterier inkluderer kokker (sferiske), baciller (stangformede) og cocco bacilli (korte, fyldige stenger). Fargeprosessen innebærer å dekke det fikserte smøret med fargeløsning i noen minutter, deretter vaske og tørke før mikroskopisk undersøkelse. Resultatene viser bakteriekulturer med spesifikke former og arrangementer.

Para llevar

  • 🔬 Enkel fargingsteknikk for bakterier.
  • 🦠 Bruk av metylblå eller fortynnet karbolfusjon.
  • 📏 Identifisering av bakteriers form og størrelse.
  • 🧪 Forberedelse av bakteriesmear er avgjørende.
  • 🔥 Varmefiksering dreper mikroorganismer.
  • 💧 Fargeprosessen tar 3-4 minutter.
  • 🔍 Mikroskopisk undersøkelse under 100x forstørrelse.
  • 📊 Resultater viser bakteriearrangementer.
  • 📚 Neste video om gramfargingsteknikk.
  • 🧼 Aseptiske forhold er viktige i laboratoriet.

Cronología

  • 00:00:00 - 00:05:00

    I denne videoen demonstreres en enkel fargemetode for bakterier, kjent som enkel farging. Denne metoden bruker kun én type farge for å visualisere bakterienes form, størrelse og arrangement. Bakterier kan ha forskjellige former, inkludert kokker (spherical), baciller (rod-shaped), og coccobacilli (short plump rods). Det diskuteres også forskjellige arrangementer av bakterier, som diplo (par), tetrader (kvadrat av fire), streptokokker (kjede), og stafylokokker (kluster).

  • 00:05:00 - 00:10:55

    Videoen fortsetter med en detaljert beskrivelse av hvordan man forbereder en bakteriesmear og utfører den enkle fargemetoden. Først må man lage en bakteriesmear ved å bruke en steril inokulasjonsløk og destillert vann. Etter å ha tørket smøret, blir det fikset med varme før farging med metylblå eller fortynnet karbolfusjon. Etter farging og skylling er smøret klart for mikroskopisk undersøkelse, hvor bakterienes form og arrangement kan observeres. Resultatene fra fargingen oppsummeres, og det nevnes at neste video vil dekke gramfarging.

Mapa mental

Vídeo de preguntas y respuestas

  • What is simple staining?

    Simple staining is a basic technique used to visualize bacteria using a single stain to identify their shape, size, and arrangement.

  • What stains are used in simple staining?

    Common stains include methylene blue and dilute carbol fusion.

  • What are the major shapes of bacteria?

    The major shapes include cocci (spherical), bacilli (rod-shaped), and cocco bacilli (short plumpy rods).

  • How is a bacterial smear prepared?

    A bacterial smear is prepared by emulsifying a small portion of bacterial culture in sterile water on a glass slide and allowing it to air dry.

  • What is heat fixation?

    Heat fixation involves passing the dried smear through a flame to kill microorganisms and fix the smear to the glass.

  • How is the staining process conducted?

    The fixed smear is covered with a staining solution for a few minutes, then washed and dried before microscopic examination.

  • What can be determined from simple staining?

    Simple staining allows for the determination of bacterial shape, size, and arrangement.

  • What is the next topic in the series?

    The next video will cover the gram staining technique, which is a differential staining method.

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  • 00:00:00
    [Music]
  • 00:00:10
    in this video demonstration
  • 00:00:11
    we will see about a bacterial staining
  • 00:00:14
    technique
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    simple staining this protocol is very
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    simple
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    and basic which helps to visualize the
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    bacteria
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    simple staining here only one stain is
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    used
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    so all bacteria are stained similarly
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    just to identify their shape size and
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    the arrangement of the bacteria
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    stains like either loafuller's methyl
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    and blue or
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    dilute carbol fusion staining solution
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    is used
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    before that we will have a brief
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    discussion on
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    the different shapes of bacteria some
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    bacteria are
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    spherical shaped this type of bacteria
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    is called as
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    cockeye and the bacteria can be
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    rod shaped this type of bacteria is
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    called as
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    bacilli if it is in between
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    that is short plumpy rod shaped
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    then they are called as cocco bacilli
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    these are the major shapes of the
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    bacteria
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    other shape of the bacteria are spiral
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    shaped
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    this type of bacteria is called as
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    spirula
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    the example of bacteria is spirulum
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    species
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    then corkscrew shaped the example
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    spirec eats curved rod shaped that is
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    comma shaped the example vibrio species
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    these are the other shapes of the
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    bacteria
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    now we will see about the different
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    arrangements observed in the bacteria
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    you know the spherical shaped bacteria
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    is called as
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    cockeye but if the two i
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    arranged in pair then it is called as
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    diplo
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    cocky if it is in square of
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    four i then it is called as deterad
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    the example of bacteria is micro caucus
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    species and it can be arranged in
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    cube of eight cockeye the example
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    sarsana
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    the cocky can be arranged in chain
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    the example of bacteria is strepto
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    caucus species
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    and the cockeye can be arranged in
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    clusters
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    the example staphylococcus species
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    these are some arrangements observed in
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    cockeye
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    you know the rod-shaped bacteria is
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    called as bacilli
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    it can be arranged as chain of bacilli
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    the example strepto bacillus species
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    and the bacilli can also have palisade
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    arrangement the example carney bacterium
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    species
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    these are some arrangements noticed in
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    bacilli
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    coming to the simple staining technique
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    before staining we need to prepare the
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    bacterial smear
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    flame the inoculation loop to red heat
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    and
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    allow it to cool
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    take a clean microscopic glass slide
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    place a drop of sterile distilled water
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    on a clean glass slide with the help of
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    the inoculation loop
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    take the bacterial culture plate and
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    open near the spirit lamp
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    take a small portion of the bacterial
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    colony
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    from the bacterial culture plate and
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    emulsify it
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    with a drop of water on the glass slide
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    after emulsifying spread the smear to
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    the size of
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    about one centimeter square so as to
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    form a thin
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    film after spreading allow it to air dry
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    it may take two to three minutes for air
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    drying
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    once the smear get dried fix the smear
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    by heat by passing this mirror through
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    the top portion
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    of the flame two to three times with a
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    slow sweeping motion
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    this is called heat fixation of the
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    smear
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    never heat the smear side avoid
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    excess heating the glass should be just
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    warm
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    to be borne by the back of the hand two
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    things
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    happens by the heat fixation one
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    the heating kills the microorganism and
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    secondly heating fixes the smear to the
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    glass
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    so that the smear will not be washed off
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    when treated with
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    staining solution the procedure for
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    the simple staining after
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    the preparation of this mirror place the
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    fixed smear
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    on the staining rack add methyl and blue
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    staining solution
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    over the smear cover the entire smear
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    with the staining solution
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    and allow it to stand for three to four
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    minutes
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    after three to four minutes wash it with
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    the distilled water
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    after washing blot dry the smear with
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    blotting paper
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    or by using tissue paper now the smear
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    is ready
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    for the microscopic examination
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    the microscopic examination
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    place the stain slide over the stage
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    examine the slide under 100x oil
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    immersion
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    objective microscopy the procedure
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    for the simple staining can also be done
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    using
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    dilute carbo fusion staining solution
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    only changes add dilute carbofusion
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    staining solution
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    over the fixed smear and allow it to
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    stand for
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    three to four minutes instead of using
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    methylene blue staining solution
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    rest of the procedure is same
  • 00:05:48
    the video demonstration first we will
  • 00:05:51
    see about the
  • 00:05:52
    preparation of the bacterial smear the
  • 00:05:55
    materials
  • 00:05:56
    required are the bacterial culture plate
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    the inoculation loop the distilled water
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    the spirit lamp and the microscopic
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    glass slide
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    these are the requirements for preparing
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    the bacterial smear
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    this experiment can be conducted over
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    the laboratory tabletop
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    with certain aseptic precautions like
  • 00:06:18
    disinfecting the table surface
  • 00:06:20
    before experiment with 70 alcohol and by
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    use of spirit lamp or bunsen burner
  • 00:06:26
    while conducting the experiment
  • 00:06:29
    first ignite the spirit lamp
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    flame the inoculation loop to red heat
  • 00:06:50
    and then allow it to cool
  • 00:06:54
    take a clean microscopic glass slide
  • 00:06:59
    place a drop of sterile distilled water
  • 00:07:02
    on the clean
  • 00:07:03
    glass slide with the help of the
  • 00:07:05
    inoculation loop
  • 00:07:11
    take a small portion of the bacterial
  • 00:07:13
    colony
  • 00:07:14
    from the bacterial culture plate and
  • 00:07:17
    emulsify it with the drop of water on
  • 00:07:20
    the glass slide
  • 00:07:22
    after emulsifying spread the smear to
  • 00:07:25
    the size
  • 00:07:26
    of about 1 centimeter square so as
  • 00:07:29
    to form a thin film
  • 00:07:36
    after spreading allow it to air dry
  • 00:07:42
    it may take two to three minutes for
  • 00:07:45
    drying
  • 00:07:46
    after air drying fix the smear by heat
  • 00:07:49
    by passing over the flame two to three
  • 00:07:51
    times with
  • 00:07:52
    a slow sweeping motion this is called
  • 00:07:55
    heat fixation of smear
  • 00:07:57
    now this heat fix smear is ready for the
  • 00:08:00
    staining
  • 00:08:02
    next the procedure for the simple
  • 00:08:05
    staining technique
  • 00:08:06
    the materials required are the heat
  • 00:08:09
    fixed smear
  • 00:08:10
    the methylene blue staining solution the
  • 00:08:12
    distilled water
  • 00:08:13
    the blotting paper and the staining rack
  • 00:08:16
    these are the requirements for
  • 00:08:18
    conducting the simple staining procedure
  • 00:08:22
    after the preparation of smear place the
  • 00:08:25
    fixed smear
  • 00:08:26
    on the staining rack
  • 00:08:29
    add the methyl and blue staining
  • 00:08:31
    solution over the smear
  • 00:08:34
    cover the entire smear with the staining
  • 00:08:36
    solution
  • 00:08:37
    and allow it to stand for three to four
  • 00:08:40
    minutes
  • 00:08:47
    after three to four minutes wash it with
  • 00:08:49
    the distilled water
  • 00:09:02
    after washing blot dry the smear with
  • 00:09:05
    the blotting paper
  • 00:09:10
    now the stain smear is ready for
  • 00:09:14
    the microscopic examination
  • 00:09:17
    the microscopic examination
  • 00:09:20
    place the stain slide over the stage
  • 00:09:23
    examine the slide under 100x oil
  • 00:09:27
    immersion
  • 00:09:27
    objective microscopy on microscopic
  • 00:09:30
    examination
  • 00:09:31
    since we have used methyl and blue
  • 00:09:34
    staining solution the microorganism
  • 00:09:37
    stains blue here the organisms are
  • 00:09:40
    spherical shape so they are cockeye
  • 00:09:44
    arrangement there are arranged in
  • 00:09:47
    clusters here
  • 00:09:49
    by simple staining method we can only
  • 00:09:51
    determine the bacterial shape
  • 00:09:53
    and their arrangement and size so
  • 00:09:56
    the result is written as the bacterial
  • 00:09:59
    culture
  • 00:10:00
    stained using the simple staining
  • 00:10:02
    technique is i
  • 00:10:03
    with cluster arrangement
  • 00:10:07
    in case if we are using dilute
  • 00:10:09
    carbo-fusion staining solution
  • 00:10:11
    the microorganism stains pink
  • 00:10:15
    here the organisms are rod shape
  • 00:10:18
    so they are bacilli arrangement
  • 00:10:21
    there is no specific arrangement noticed
  • 00:10:25
    so the result is written as the
  • 00:10:27
    bacterial culture
  • 00:10:28
    stained using the simple staining
  • 00:10:30
    technique is bacilli
  • 00:10:32
    with no specific arrangement
  • 00:10:36
    with this we are coming to the end of
  • 00:10:38
    the simple staining technique
  • 00:10:40
    in next video demonstration we will see
  • 00:10:42
    about the gram staining technique
  • 00:10:44
    which is a differential staining hope
  • 00:10:47
    the demonstration
  • 00:10:48
    is informative and useful thank you
Etiquetas
  • bacterial staining
  • simple staining
  • bacterial shapes
  • heat fixation
  • methylene blue
  • carbol fusion
  • microscopic examination
  • bacterial arrangements
  • laboratory techniques
  • gram staining