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NEUROTRAILBLAZERS · TEACHING MATERIALS

Glia

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Contents · 18 slides
  1. 1. Glia
  2. 2. Session outcomes (60 minutes)
  3. 3. Pedagogical arc
  4. 4. Why this matters technically
  5. 5. Orientation: the stake
  6. 6. Astrocyte morphology in a synaptic neighbourhood
  7. 7. Real data: an astrocytic process
  8. 8. Real data: cortical vasculature
  9. 9. Microglia cues
  10. 10. Oligodendrocyte-associated cue context
  11. 11. Myelin-context interpretation
  12. 12. Operational glia triage protocol
  13. 13. Metrics to track
  14. 14. Misconceptions to correct
  15. 15. Activity
  16. 16. Rubric checkpoint
  17. 17. The three classes, schematically
  18. 18. References and attribution

Slide 1 of 18

H01 electron microscopy with object segmentation and original 2 µm scale bar

Human cortex · H01
Object segmentation over electron microscopy

H01 release · Lichtman Lab / Harvard & Connectomics at Google · CC BY 4.0
Shapson-Coe et al. (2024) · doi:10.1126/science.adk4858

Technical Course · Unit 07

Glia

Astrocytes, oligodendrocytes and microglia in EM — and why a single glia–neuron merge is one of the most expensive errors in the volume.

Slide 2 of 18

Session outcomes (60 minutes)

Slide 3 of 18

Pedagogical arc

Slide 4 of 18

Why this matters technically

Figure description and teaching notes

Walk one merge (unit §1): a fine astrocytic process fused onto a dendrite. The neuron gains a branch that does not exist. Because astrocytic processes ensheathe synapses, the merged path runs directly past many of them, and those detections get attributed to the neuron — inputs from cells it never contacted. The false inputs are spatially local, so they inflate local clustering and short-range connectivity.
Instructor script: "Again the bias points toward an interesting result. That is why glia–neuron merges rank above many neuron–neuron splits in the queue, even though the split is more visually obvious."

Slide 5 of 18

Orientation: the stake

Glia occupy roughly 20–40% of cortical volume.

Source: Pat Rivlin training materials (MICrONS proofreading deck), RIV-GLIA S01.

Figure description and teaching notes

Set the stake before anyone starts looking: a glia–neuron merge does not merely add a branch — it drags a neuron's arbor past synapses it never contacted, so the false-input yield per micrometre of merged path is unusually high.
Each glial class has one near-diagnostic feature. Learn those three first: astrocyte — glycogen granules; oligodendrocyte — the darkest nucleus in the field; microglia — a dense elongated nucleus plus lysosomal content.

Slide 6 of 18

Astrocyte morphology in a synaptic neighbourhood

An astrocytic process is space-filling; a neurite is a tube.

Source: Ventura RE, "Astrocytes," SynapseWeb (Kristen M. Harris Lab, UT Austin), synapseweb.clm.utexas.edu/astrocytes. © The University of Texas at Austin; no reuse licence is stated — confirm permission before this deck is shared outside the course.

Figure description and teaching notes

Read cross-sectional shape before anything else. An astrocytic process takes whatever contour is left over after the neurites pack; a neurite holds a tube shape of its own. That cue survives weak staining better than glycogen granules do, which makes it the one to reach for where everything else fails.
Other astrocyte cues from unit §2: pale "watery" cytoplasm with few organelles; glycogen granules 20–30 nm, very dark, clustered (neurons do not contain them); perisynaptic processes under 100 nm thick; endfeet flattened against vessels; no vesicles, no PSDs, few microtubules.

Slide 7 of 18

Real data: an astrocytic process

  • Pale cytoplasm, irregular space-filling outline that wraps its neighbours.
  • Labelled 'astrocyte' by H01's subcompartment model.

Source: H01 release (Lichtman Lab, Harvard; Connectomics at Google), CC BY 4.0 · Shapson-Coe et al. 2024, doi:10.1126/science.adk4858 · site render.

Figure description and teaching notes

Instructor script: "Trace the outline with your finger. Does it hold its own shape, or does it fill the gaps the neurites left?"
Use the unit's check-yourself: a pale, irregular profile between three neurites, no glycogen visible. Probably astrocytic — medium confidence at best. Absence of glycogen is weak evidence because granules are clustered and sparse. Better evidence, in order: space-filling behaviour over several sections; no vesicles or PSD on it; continuity to an endfoot; microtubules at higher magnification.

Slide 8 of 18

Real data: cortical vasculature

  • H01 contains ~230 mm of blood vessels across ~1 mm³.
  • Vessels are a common site of merge errors — glia wrap them closely.

Source: H01 release (Lichtman Lab, Harvard; Connectomics at Google), CC BY 4.0 · Shapson-Coe et al. 2024, doi:10.1126/science.adk4858 · site render.

Figure description and teaching notes

Walk the unit's worked example here: a pale profile a few hundred nanometres thick, pressed flat against a capillary, currently assigned to a nearby dendrite. Rule out synaptic participation first — even when the vascular context is shouting the answer. Glycogen absent: weak evidence, noted not spent. Over 5–10 sections the contour spreads to track the vessel wall: space-filling. Context: a flattened expansion covering a vessel is an endfoot. Dismiss microglia for cause — the parent process is pale and organelle-poor, not inclusion-rich.
Call: astrocytic endfoot, high confidence; the dendrite assignment is a glia–neuron merge and goes above conspicuous splits in the queue.

Slide 9 of 18

Microglia cues

Weight nuclear evidence above process shape.

Source: Pat Rivlin training materials (MICrONS proofreading deck), RIV-GLIA S09.

Figure description and teaching notes

Look for the pairing that carries the call: a dark, often elongated or bean-shaped nucleus with heterochromatin clumped against the envelope, plus lysosomal and phagosomal content in dense cytoplasm with long, narrow ER cisternae. Microglia are the least stereotyped of the three classes because morphology tracks activation state — which is why nuclear evidence outranks process shape, and why a microglia call needs a nucleus or a lysosome-rich parent process, never a fragment.

Slide 10 of 18

Oligodendrocyte-associated cue context

No extracted micrograph exists for this class; the full cue table is in Unit 07 §2.

Figure description and teaching notes

Instructor script: "If you remember one thing about oligodendrocytes, it is the nucleus: distinctly darker than neuronal or astrocytic nuclei, usually identifiable at a glance." Then spend the time on the OPC: the correct action for a "not quite" cell is to flag rather than force. A flagged OPC candidate is a useful annotation; a forced wrong class is not.

Slide 11 of 18

Myelin-context interpretation

  • Myelin is the fastest route to an oligodendrocyte identification: follow a sheath back to the soma that owns it.
  • A merge across a myelinated axon costs more than one across an unmyelinated process: the axon it corrupts is long-range.

Source: H01 release (Lichtman Lab, Harvard; Connectomics at Google), CC BY 4.0 · Shapson-Coe et al. 2024, doi:10.1126/science.adk4858 · site render.

Figure description and teaching notes

The panel highlights every myelinated axon the H01 myelin mask finds in one grey-matter field — a small minority of profiles in cortex. Instructor cue: ask how myelin context changes proofreading priority. Context cues from the protocol: the inner or outer tongue of a myelin sheath, or a paranodal loop, is a high-confidence oligodendrocyte call.

Slide 12 of 18

Operational glia triage protocol

  1. Identify likely class from morphology/context.
  2. Validate local boundary integrity.
  3. Estimate downstream risk if left uncorrected.
  4. Route to immediate correction or adjudication queue.
Figure description and teaching notes

The unit's identification protocol, which feeds step 1: (1) soma? read the nucleus first — very dark and round -> oligodendrocyte; dark, elongated, peripheral heterochromatin plus lysosomes -> microglia; pale, irregular, glycogen -> astrocyte; pale but "not quite" -> OPC candidate, flag. (2) Synaptic participation -> neurite, go to Unit 06. (3) Glycogen -> astrocyte. (4) Cross-sectional character over 5–10 sections. (5) Context: endfoot, myelin tongue, lysosome-rich parent. (6) Unresolved -> uncertain, with the missing cue named.
Context is the strongest cue in the protocol, but it is only safe after the exclusion steps have run.

Slide 13 of 18

Metrics to track

Slide 14 of 18

Misconceptions to correct

Slide 15 of 18

Activity

Classify two ambiguous glia-neuron interfaces and submit:

Slide 16 of 18

Rubric checkpoint

Slide 17 of 18

The three classes, schematically

Source: Holly Fischer, Wikimedia Commons, File:Glial_Cell_Types.png. CC BY 3.0 Unported.

Slide 18 of 18

References and attribution