Domain: Healthcare — Basic Immunology
Target concept: Hemolysis Control Experiment

Video generation prompt: Prepare antigen (sheep red blood cells), antibody (anti-sheep red blood cell serum), and complement (fresh guinea pig serum). Add, on the left side, the antigen:antibody:complement in a volume ratio of 2:1:2 into a sterile tube, gently mix, and incubate at 37°C in a water bath for 30 minutes. On the right side, set up a complement-free control group (antigen + antibody) and an antibody-free control group (antigen + complement), and incubate them simultaneously. Display the two groups side by side.

Evaluation guide:
(i) Target concept: Complement-dependent hemolysis is a binary, tripartite requirement: antigen, specific antibody, and active complement must all be present simultaneously to lyse sheep RBCs.
Minimum mechanism: Antibody binds SRBCs → forms immune complexes → activates classical complement pathway → membrane attack complex (MAC) forms → osmotic lysis → release of hemoglobin → supernatant clears and turns pale pink.
(ii)Storyline: Frame 1: Three reagents dispensed into left tube (visually distinct colors/volumes); two control pairs placed right-side adjacent. Frame 2: Gentle mixing applied only to left tube (visible agitation, no splashing). Frame 3: All tubes placed side-by-side in water bath (thermal context implied). Frame 4: Post-incubation state: left tube shows clear supernatant + pellet/ghost cells; both right tubes retain uniform opaque red suspension. Perspective: fixed frontal, eye-level view; full-tube visibility maintained; no occlusion; depth preserved via consistent tube alignment and shared background.

Manual:
Experimental Procedure
1. Place antigen (sheep RBCs), antibody (anti-sheep serum), and complement (fresh guinea pig serum) into left sterile tube in 2:1:2 volume ratio.
2. Place antigen+antibody in one right tube; antigen+complement in second right tube.
3. Gently mix only left tube; incubate all tubes at 37°C for 30 min (implied by water bath).
Observation of Experimental Phenomena
Left tube: clear/pale supernatant (hemolysis). Right antigen+antibody tube: opaque red (no lysis). Right antigen+complement tube: opaque red (no lysis).
Summary of Experimental Principles
Hemolysis requires antigen-antibody complex formation AND functional complement activation — only the triple combination yields lysis.

Prompt Grounding rubric:
level_1: No tubes, reagents, or water bath appear at all; or none of the three reagents (SRBC suspension, anti-SRBC antiserum, guinea-pig serum) is identifiable in any tube; the left (three-component) tube and the two right-side control tubes are never shown together in the same shot.
level_2: At least one tube with a reagent appears, but none of the reagents is identifiable as antigen, antibody, or complement (no legible label or clearly distinct liquid); the left tube and the two right-side tubes are never framed together; there's no visible incubation setup; nothing distinguishes the three-component tube from either two-component control.
level_3: Three tubes are visible together in one shot: the left tube receives antigen, antibody, and complement; one right-side tube receives antigen + antibody only; the other right-side tube receives antigen + complement only; the three reagents are visually distinguishable by color or label; every tube has the right components in it even though mixing/incubation isn't shown yet.
level_4: Everything at the 3-point level holds, and in addition a gentle mixing motion is shown only for the left tube (neither right-side tube is stirred); a water bath appears behind all three tubes with a thermometer or digital readout showing roughly 37°C (no steam or condensation, since 37°C would not produce visible steam); no reagent ends up in the wrong tube.
level_5: Everything at the 4-point level holds, and in addition the left tube visibly receives its three additions as separate, comparably sized pours (the middle one, antibody, clearly smaller than the other two, roughly matching a 2:1:2 feel) rather than one combined pour; each right-side tube's antigen portion looks the same size as the left tube's antigen portion; all labels ('SRBC', 'Anti-SRBC', 'GPS' or equivalent) stay legible; the water bath's thermometer/readout stays visible and steady; the three tubes stay in a straight, parallel row with no rotation or depth shift between them.

Scientific and Causal Correctness rubric:
level_1: None of the three tubes shows any color separation; all three look like uniform opaque red suspensions from start to finish, or a color change happens with no visible tie to which reagents are in which tube.
level_2: A color change happens, but it shows up in the wrong tube(s), e.g., one of the two-component control tubes clears up, or the three-component left tube stays fully opaque while a control tube clears, or every tube changes together with no difference between them.
level_3: By the end, the left tube (all three components) shows a see-through or pale pink upper layer over a settled residue, while both right-side tubes (each missing one component) stay uniformly cloudy dark red with no clearing; this pattern holds steadily once it appears, in only the left tube.
level_4: Everything at the 3-point level holds, and in addition the clearing in the left tube is complete, not partial (no cloudy patches remaining in its upper portion); neither right-side control tube shows even slight lightening or a two-layer look; once the left tube's clear layer appears it doesn't turn cloudy again, and no color spreads from the left tube into either control tube.
level_5: Everything at the 4-point level holds, and in addition the left tube ends with a sharp, clean boundary between a clear/pale top layer and a settled reddish residue at the bottom; both control tubes remain a single, even, opaque dark red from top to bottom with no boundary at all; all three tubes look like the same shade of red before the incubation step, so the later difference is attributable to the reagent combination and not to a starting-color mismatch.

Spatiotemporal Consistency rubric:
level_1: One or more tubes jump to a new spot or angle between shots with no visible movement causing it; a reagent (SRBC suspension, antiserum, or guinea-pig serum) appears in or vanishes from a tube with no pour or drop shown; the left tube's mixing action starts with the liquid already swirling before any stirring tool or hand touches it; the three-tube arrangement cuts from a close-up to a wide shot with no transition; or the lighting around the tubes flips abruptly between very different brightness levels with no cause.
level_2: The left tube or either right-side control tube shifts position or tilt partway through the clip; the mixing tool (pipette tip, stirring rod, or vortexer) touches the left tube but the liquid inside doesn't move in response, or moves before contact; the row of tubes changes order (e.g., left and right swap) between frames; or the lighting on the left tube looks noticeably different in brightness from the lighting on the right-side tubes despite being filmed in the same continuous shot.
level_3: The left tube and the two right-side control tubes stay in the same spot and orientation for the whole clip; the mixing action on the left tube is a single continuous stirring or swirling motion that clearly starts from a tool or hand entering the frame and touching the liquid; the right-side tubes never move or get touched during this mixing step; none of the three tubes changes shape or suddenly warps.
level_4: Everything at the 3-point level holds, and in addition the three tubes keep the same spacing and height relative to each other throughout; the left tube's mixing motion finishes and the tool withdraws before any change appears in either right-side tube; each tube's liquid surface stays flat and still whenever no mixing or pouring is happening in that tube; no tube's edge or label flickers or blurs between adjacent frames.
level_5: Everything at the 4-point level holds, and in addition the left tube's mixing motion speeds up and slows down the way a hand or vortexer naturally would rather than moving at one constant speed; ripples caused by the stirring spread outward across the left tube's liquid and settle down afterward; the boundary between the settled solid and the clear liquid in the left tube (once it appears) stays sharp and doesn't blur or flicker from one frame to the next; the same side of the frame keeps the same tube throughout (no left/right relabeling).