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<span style="font-size:90%">'''Navigation:''' [[MalariaETC Index|Return to Main Malaria Index]]''</span>
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<span style="font-size:90%">A thick film is prepared by placing a small drop of blood on a slide then spreading it in a circular motion. The thick layer acheived is then air-dried without fixation.
</br></br>'''The principles are:'''</br>
*<span style="font-size:90%">The blood layer will be many layers thick (varying from 6-20 accross the specimen)
*<span style="font-size:90%">The erythrocytes are unfixed, so will be lysed during staining appearing only as debris.
*<span style="font-size:90%">The Giemsa stain will stain and distinguish the remaining white cells and parasites.
*<span style="font-size:90%">This concentration effect allows parasites to be detected with high sensitivity
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<span style="font-size:90%">A thick film is prepared by placing a small drop of blood on a slide then spreading it in a circular motion. The thick layer acheived is then air-dried without fixation.</br></br>The principles are:</br>
Typical appearances of a case of ''P.falciparum'' with easily detected trophozoites are shown below.</br></br>
*<span style="font-size:90%">The blood will therefore be many layers thick (around 6-20) compared with the single layer of a thin film
*<span style="font-size:90%">The erythrocytes are unfixed so will be lysed during staining appearning only as debris.
*<span style="font-size:90%">The Giemsa stain will therefore stain and distinguish the remaining white cells, parasites.
*<span style="font-size:90%">This allows parasites to be detected with high sensitivity using fewer microscopic fields
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<span style="font-size:90%">All these features allow thick films to be highly sensitive for parasite detection, but also introduce difficulties for morphologists. Principally staining inconsistencies as stain variable penetrates the thick cell layer and disruption of parasites caused by drying and shrinkage, finally the technique disrupts or eliminates red cells that provide valuable clues to species. Many skills are transferable from thin to thick film specimens but the differences require microscopists  to be aware of potential artefact and to have experience of thick blood films. Typical appearances of a case of ''P.falciparum'' with easily detected trophozoites are shown below.</br></br>
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File:TF1b.jpg|<span style="font-size:80%">Low magnification view for scale, 3 regions marked</span>|link={{filepath:TF1b.jpg}}
File:TF1b.jpg|<span style="font-size:80%">Low magnification view for scale, 3 regions marked</span>|link={{filepath:TF1b.jpg}}
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File:TF1e.jpg|<span style="font-size:80%">Region C: 2 trophozoites in one group</span>|link={{filepath:TF1e.jpg}}</gallery>"
File:TF1e.jpg|<span style="font-size:80%">Region C: 2 trophozoites in one group</span>|link={{filepath:TF1e.jpg}}</gallery>"
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Note the differences in recognition - the typical ring form and vacuole of the parasite are not as easy to distinguish and chromatin dots may appear to separate from parasite cytoplasm while the absence of intact red cells takes away important clues to parasite size and distribution. Some strengths and weaknesses of each approach are summarised in the table below:</br>
<span style="font-size:90%">Note the differences in recognition - the typical ring form and vacuole of the parasite are not as easy to distinguish and chromatin dots may appear to separate from parasite cytoplasm while the absence of intact red cells takes away important clues to parasite size, distribution within the red cell, and any red cell changes. This is illustrated in the image below</span>


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File:2_Trophs_MP.jpg|<span style="font-size:80%">Two ring frms of ''P.falciparum''</span>|link={{filepath:2_Trophs_MP.jpg}}
File:3_Trophs_HP.jpg|<span style="font-size:80%">High-power image of the parasites</span>|link={{filepath:3_Trophs_HP.jpg}}
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'''Detailed Sections:'''</br></br>


<div style="width: 95%; font-size:90%;">  
'''Section 1: strengths and weaknesses of thick malaria films''' a comparison different morphological approaches to malaria diagnosis.
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</br><span style="font-size:200%">&#x2192;</span> [[Thick_vs_Thin_film_comparison|Click to see comparison of thick and thin film approaches]]</br></br>
!colspan="3" style = "background:#e1f1fd; border:solid; border-width: 3px;"|<span style="font-size:90%;">'''COMPARISON OF THICK AND THIN FILMS'''</span></br>
'''Section 2: recognising parasites on thick films''' Initial assessment - recognising staining debris and identifying parasites.
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</br><span style="font-size:200%">&#x2192;</span> [[Thick films - parasites and debris|Click to see the approach to identify parasites and distinguish debris]]</br></br>
! Feature !! Thick Film !! Thin Film
'''Section 3: Species identification on thick films''' - the possibilities and limitations of species identification.
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</br><span style="font-size:200%">&#x2192;</span> [[Thick films - parasites identification|Click to see examples of malaria species, stage or pigment on thick films]]</br></br>
| Sensitivity for detection || Higher: detects low parasitaemia ~5–10 parasites/µL || Lower: generally needs ~50 parasites/µL for reliable detection)
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| Species Identification || Poor: RBC morphology lost and species-specific features may be difficult || Excellent: Parasite morphology and RBC characteristics are readily observed
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| Quantification of parasitaemia || Difficult: requires estimation so is imprecise || Easier: parasites can be counted per number of RBCs
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| Preparation and staining || Longer: requires air drying before careful staining to avoid artefact || Faster: films are fixed and stained immediately with clearer morphology
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Latest revision as of 10:00, 27 February 2025


Navigation: Return to Main Malaria Index



OVERVIEW OF THICK FILMS


A thick film is prepared by placing a small drop of blood on a slide then spreading it in a circular motion. The thick layer acheived is then air-dried without fixation.

The principles are:

  • The blood layer will be many layers thick (varying from 6-20 accross the specimen)
  • The erythrocytes are unfixed, so will be lysed during staining appearing only as debris.
  • The Giemsa stain will stain and distinguish the remaining white cells and parasites.
  • This concentration effect allows parasites to be detected with high sensitivity


Typical appearances of a case of P.falciparum with easily detected trophozoites are shown below.

"


Note the differences in recognition - the typical ring form and vacuole of the parasite are not as easy to distinguish and chromatin dots may appear to separate from parasite cytoplasm while the absence of intact red cells takes away important clues to parasite size, distribution within the red cell, and any red cell changes. This is illustrated in the image below



Detailed Sections:

Section 1: strengths and weaknesses of thick malaria films a comparison different morphological approaches to malaria diagnosis.
Click to see comparison of thick and thin film approaches

Section 2: recognising parasites on thick films Initial assessment - recognising staining debris and identifying parasites.
Click to see the approach to identify parasites and distinguish debris

Section 3: Species identification on thick films - the possibilities and limitations of species identification.
Click to see examples of malaria species, stage or pigment on thick films