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Edited training material "Automated Cell Annotation" upto slide 10 #5604

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Expand Up @@ -36,11 +36,11 @@

### What is Cell Annotation?

- Categorising cells into cell type categories based on transcriptomic data
- Classify cells in your data into different cell types based on gene expression data

- Can be done manually or automatically

- Is possible due to single-cell sequencing technology
- Single-cell sequencing technology provides higher resolution than bulk RNA-seq
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I feel like this point is missing something to explicitly link it to the cell annotation - maybe combine it with the start of the old version like 'Is possible because single-cell seq tech provides higher res...'?


![Basic pipeline for automated cell annotation](../../images/scrna-cell-annotation/cell-annotation.png)

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---

### Why is it Important?
### Why is Cell Annotation Important?

- Able to process and analyse single cell data much faster than manual analysis

- To understand the composition of cell types in samples (cellular heterogeneity)

- To compare changes in cell populations or states across different condition and phenotypes
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Suggested change
- To compare changes in cell populations or states across different condition and phenotypes
- To compare changes in cell populations or states across different conditions and phenotypes


- To perform differential expression within each cell type to avoid signal dilution from mixed cell type population

- To identify novel cell states and study some cell population further

???

The power of single-cell RNA-seq lies in its ability to capture the transcriptome at single-cell resolution.

However, a significant challenge is accurately classifying cells into distinct types before beginning downstream analysis.

Once cells are annotated, we can examine the composition of cell types within each sample and compare them across conditions.

This precise classification enables differential expression analysis within specific cell types, a key objective of single-cell experiments.


---

### Why Automate Cell Annotation?

- Each single-cell experiment can generate data for thousands of cells

- Manual annotation is time consuming and requires domain expertise

- Can produce results more consistently allowing for reproducibility of results

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- Cell annotation uses sc-RNA seq data

- Gene expressions are stored in a gene expression matrix (X)
- Gene expressions are stored in a gene expression matrix (X) <br><br>

Common data types:

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---

### Challenges
### Challenges in Automated Cell Type Annotation

- Noise due to amplification techniques, varying sequencing depths, and sequencing errors

- Data is often noisy due to amplification techniques, varying sequencing depths, and errors in the reads
- Difference in QC steps for reference and query dataset may introduce a bias

- Cell type definitions are inherently subjective and may be suboptimal

- Lack of a suitable reference panel for the query dataset can result in inaccurate classification, especially when dealing with unknown cell states

- Dealing with unknown cell types due to undiscovered classifications


???

There are some challenges that need to be faced in order to perform automated cell annotation:
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### Manual Cell Annotation

- Generate a cluster map and manually annotate each clusters
- Requires known marker genes for cell types of interest

- Generate the UMAP/tSNE (see below) to visualize the expression values

- Requires the researchers to find known marker genes in the data
- If you have clustered the cells, you can use dotplots or violin plots to measure the average expression of these genes

![Various cell cluster diagrams showing the expression values of various marker genes](../../images/scrna-cell-annotation/manual-annotation.png)

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.pull-left[

- The process of identifying unique gene expressions that can be used for identifying cell types
- The process of identifying genes that are uniquely expressed in certain cell types

- Genes are selected based on how differently they are expressed across different cell types

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