Domain: Healthcare — Basic Immunology
Target concept: Microcytic, Hypochromic Red Blood Cell Morphology in Iron Deficiency Anemia

Video generation prompt: Collect peripheral blood from patients with iron deficiency anemia, prepare a blood smear, and stain with Wright-Giemsa. Observe the morphology of red blood cells under oil immersion microscopy after drying.

Evaluation guide:
(i) Target concept: Microscopic identification of microcytic, hypochromic red blood cells in peripheral blood smear from iron deficiency anemia patient.
Minimum mechanism: Blood must be freshly collected, smeared as a monolayer, properly stained with Wright-Giemsa (requiring both eosin and methylene blue components), fully air-dried to fix morphology, and examined under oil immersion to resolve biconcave disc shape and central pallor.
(ii)Storyline: [Phase 1: Collection] Lancet contacts fingertip → blood emerges as cohesive droplet. [Phase 2: Smear] Droplet contacts slide → spreads unidirectionally into tapered monolayer with feathered edge. [Phase 3: Staining] Stain solution floods smear → covers entire thin area → develops biphasic color separation. [Phase 4: Drying & Oil] Slide surface transitions from wet-shiny to dry-matte → oil droplet placed centrally on dried smear. [Phase 5: Observation] Microscope field darkens → focuses smoothly → resolves high-magnification field containing exclusively small, pale-centered RBCs against clean background; all phases occur from consistent lateral or overhead perspective with no occlusion of critical interfaces (blood/slide, stain/smear, oil/lens).

Manual:
Experimental Procedure
1. Prick fingertip with lancet; collect drop of peripheral blood.
2. Transfer blood to clean glass slide; spread into thin, even monolayer smear.
3. Air-dry smear completely.
4. Flood smear with Wright-Giemsa stain; incubate 5–10 min.
5. Rinse gently; air-dry again.
6. Apply immersion oil; observe under 100× oil immersion objective.
Observation of Experimental Phenomena
Microscopic field shows numerous red blood cells that are smaller than white blood cells and exhibit pale central areas (hypochromia) with sharp borders.
Summary of Experimental Principles
Iron deficiency anemia causes impaired heme synthesis → reduced hemoglobin content per RBC → microcytosis and hypochromia; Wright-Giemsa stain differentially colors hemoglobin (pink-orange) and cytoplasm/nuclei (blue-purple), enabling visualization of central pallor and size reduction.

Prompt Grounding rubric:
level_1: No blood collection, no smear preparation, no staining, no microscope view; none of the four required objects (capillary tube/blood lancet, glass slide, Wright-Giemsa stain bottle, oil immersion microscope) appear; no red blood cell morphology shown.
level_2: At least one object appears (e.g., glass slide or microscope), but zero steps are performed in correct sequence; no visible blood sample, no stain application, no oil immersion field; no red blood cells present or identifiable.
level_3: All four core objects appear: capillary tube or lancet, clean glass slide, Wright-Giemsa stain container, and oil immersion microscope; blood is visibly placed on slide, stain is applied to smear area, and final frame shows a microscopic field containing red blood cells, but sequence is disordered (e.g., stain applied before blood placement) or key action is missing (e.g., no spreading motion, no oil drop visible before microscopy).
level_4: All four core objects appear; blood is placed on slide, then spread into thin film, then stained with Wright-Giemsa, then dried, then oil is applied, then viewed under oil immersion; all actions are visually present and occur in correct sequential order; red blood cells are visible in final field, but one non-core flaw exists (e.g., stain container label illegible, slide edge not aligned in view, oil droplet absent from final frame though microscope is correctly set for oil immersion).
level_5: All four core objects appear with correct labeling/identity (capillary tube or lancet with skin puncture site visible, clean unscratched glass slide, Wright-Giemsa stain bottle with correct blue-red hue, oil immersion microscope with 100× objective engaged); blood is placed → spread into monolayer smear → stained → air-dried → oil applied → viewed under oil immersion; final frame shows a sharply focused, centered field containing numerous red blood cells exhibiting microcytic, hypochromic morphology (small size, pale central area, crisp margins); all actions are continuous, spatially coherent, and temporally ordered without omission or substitution.

Scientific and Causal Correctness rubric:
level_1: No scientifically valid red blood cell morphology shown; cells appear normocytic/normochromic, giant, fragmented, or absent; stain color is monochrome (e.g., only blue or only pink), or no differential staining contrast; no evidence of hypochromia or microcytosis.
level_2: Red blood cells appear, but all are uniformly large and fully hemoglobinized (no pallor, no size reduction); stain shows no biphasic blue (nuclei/cytoplasm basophilia) and pink (hemoglobin/eosinophilia) contrast; no central pallor visible; no morphological abnormality consistent with iron deficiency.
level_3: Red blood cells are present in final oil immersion field and show *some* cells with central pallor and reduced diameter relative to white blood cells or platelets; stain displays basic biphasic hue (blue-purple nuclei, pink cytoplasm), but pallor is inconsistent across cells, or microcytosis is only weakly suggested without clear size contrast; causal chain is partially supported (staining occurred, microscopy used) but morphology does not robustly reflect iron deficiency anemia.
level_4: Final field contains multiple red blood cells that are *consistently smaller than adjacent white blood cells* and exhibit *clear, well-demarcated central pallor occupying >1/3 of cell diameter*; stain shows correct Wright-Giemsa biphasic contrast (blue-purple granules/nuclei, pink-orange hemoglobin rim); all morphological features match textbook iron deficiency anemia; no contradictory features (e.g., target cells, schistocytes, polychromasia) present, but one minor non-morphological inconsistency exists (e.g., background debris misinterpreted as platelets, or slight over-staining at smear edge not affecting RBC assessment).
level_5: Final oil immersion field shows *only* microcytic, hypochromic red blood cells: every visible RBC has *uniformly reduced diameter* (visibly smaller than lymphocyte nucleus), *sharp-bordered central pallor occupying 40–60% of cell area*, and *thin, crisp cell membrane*; no normocytic, macrocytic, or hyperchromic cells present; stain demonstrates precise Wright-Giemsa differentiation: hemoglobin zone pink-orange, cytoplasm light blue, nuclei purple-blue, platelets purple; no artifacts (e.g., drying streaks, stain precipitate, oil bubbles obscuring RBCs); morphology is unambiguously diagnostic of iron deficiency anemia with zero contradictory visual evidence.

Spatiotemporal Consistency rubric:
level_1: Severe spatiotemporal rupture: objects teleport (e.g., slide appears fully stained without transition), microscope view jumps abruptly from empty field to final RBC image with no focusing motion, blood appears spontaneously on slide without contact, or lighting changes violently between frames.
level_2: Multiple discontinuities: blood placement and smear spreading occur in separate isolated shots with mismatched slide orientation; stain bottle appears in one angle, disappears, reappears in different hand; oil drop vanishes between application and microscopy; microscope focus shifts without smooth adjustment; slide rotates or flips between steps.
level_3: All steps occur within a single fixed viewpoint or logically panned sequence; objects maintain consistent spatial relationships (e.g., lancet always near finger, slide always on flat surface, microscope always stationary); no teleportation or rotation; but minor inconsistencies exist: brief cut causing temporal jump (e.g., smear appears pre-spread), or lighting flicker during stain application, or slide shifts slightly between drying and oil application without repositioning.
level_4: Continuous shot or seamlessly edited sequence with stable viewpoint; all objects retain fixed relative positions (lancet → finger → blood → slide → stain → microscope); smear spreads smoothly from thick to thin edge; stain flows evenly across smear; drying is implied by time lapse or matte finish; oil drop appears and remains intact until microscopy; microscope focus adjusts gradually to reveal RBCs; only one minor flaw: e.g., brief lens flare obscuring first 0.5s of final field, or slide edge momentarily exits frame during spreading but re-enters with same orientation.
level_5: Single unbroken take or perfectly matched cuts with identical perspective, scale, and lighting; lancet contacts fingertip → blood emerges → transfers to slide → is spread in single directional motion → stain is dropped and evenly covers smear → slide dries with uniform matte sheen → oil drop is placed centrally → microscope stage advances smoothly → focus dial turns continuously → final field resolves with zero jitter, zero occlusion, zero parallax shift; all objects maintain exact same spatial configuration and lighting throughout.