Domain: Healthcare — Basic Immunology
Target concept: IgM Detection (Simplified ELISA)

Video generation prompt: In microtiter plates pre-coated with antigen, diluted serum was added using a pipette, followed by a brief incubation and washing. Subsequently, enzyme-labeled anti-IgM antibody was added, and the plates were incubated and washed again. Finally, a chromogenic substrate was added, and the reaction was terminated.

Evaluation guide:
(i) Target concept: Enzyme-linked immunosorbent assay for IgM detection relies on antigen capture, IgM binding, enzyme-antibody conjugate recognition, and chromogenic signal amplification. Minimum mechanism: Color development requires sequential presence of antigen-coated solid phase, patient IgM, enzyme-labeled anti-IgM, and enzymatically cleavable substrate, with each step physically enabling the next; absence of any component must prevent color. (ii)Storyline: [Start] Fixed overhead view of 96-well plate with labeled antigen coating → [Event 1] Pipette draws serum from vial and dispenses into selected wells → [Event 2] Plate rests motionless (incubation implied) → [Event 3] Wash bottle rinses wells with visible fluid flow → [Event 4] Pipette adds anti-IgM to same wells → [Event 5] Second rest → [Event 6] Second wash → [Event 7] Pipette adds clear substrate → [Event 8] Wells develop uniform blue color only where prior reagents were added → [Event 9] Stop solution added → [End] Blue color stabilizes instantly. Visibility constraints: All actions must occur within static frame; no cutaways; no zooms; pipette tip and well interiors must remain fully visible at all times; color change must be discernible against white plate background.

Manual:
1. Experimental Procedure: Pipette diluted serum into antigen-coated microtiter wells → brief incubation → wash → add enzyme-labeled anti-IgM antibody → incubate → wash → add chromogenic substrate → stop reaction.
2. Observation of Experimental Phenomena: Color change from colorless to blue in wells containing IgM-positive serum; no color change in negative/control wells.
3. Summary of Experimental Principles: IgM detection via simplified ELISA: antigen-bound IgM in sample is recognized by enzyme-conjugated anti-IgM; enzymatic conversion of substrate yields visible color only when IgM is present.

Prompt Grounding rubric:
level_1: None of the following appear at all: the antigen-coated 96-well plate, the pipette, the serum vial, the anti-IgM-HRP vial, the TMB substrate vial, or any liquid being transferred into a well.
level_2: At least one of the five core objects appears (e.g., only the plate or only the pipette), but zero correct sequential actions occur: no serum is pipetted into any well, no anti-IgM is added after a wash, and no substrate is ever added.
level_3: All five core objects appear (96-well plate, pipette, serum vial, anti-IgM-HRP vial, TMB substrate vial) and at least three of the eight ordered events occur in the correct order (serum pipetted into wells, anti-IgM added, substrate added), but one or more of the two wash steps are missing, out of place, or the incubation pauses between additions are absent.
level_4: All five core objects appear and all eight ordered events occur in the correct sequence: (1) serum pipetted into wells, (2) first incubation pause (plate held motionless for at least a second), (3) first wash (visible rinse fluid moving over the wells), (4) anti-IgM-HRP pipetted into the same wells, (5) second incubation pause (again held motionless for at least a second), (6) second wash, (7) substrate pipetted in, (8) stop solution added, but one non-core flaw remains, such as the pipette tip briefly touching a well rim during a transfer, or the anti-IgM-HRP and TMB substrate vials looking so similar in shape, cap color, and bench position that a viewer could not tell them apart without reading tiny printed text.
level_5: All five core objects are present and stay clearly distinguishable from one another throughout by consistent shape, cap color, and fixed bench position (never by having to read printed vial text), and all eight ordered events (serum pipetting, first incubation pause, first wash, anti-IgM pipetting, second incubation pause, second wash, substrate pipetting, stop-solution addition) occur in strict order with no extraneous actions, no skipped steps, no object substitutions, and no steps performed out of sequence.

Scientific and Causal Correctness rubric:
level_1: No color change occurs in any well at any point; or blue appears in a well before TMB substrate has been added to it; or blue appears before anti-IgM-HRP has been added to that well.
level_2: A color change occurs but with the wrong cause: e.g., every well on the plate turns blue at the same rate regardless of whether it ever received serum; or a well turns blue immediately after serum is pipetted in, before anti-IgM-HRP or substrate are ever added; or a single well shows both blue and an unrelated color (red or yellow) with no explanation.
level_3: Blue develops only after TMB substrate is pipetted in, and only in wells shown earlier receiving both serum and anti-IgM-HRP; the control well, defined as a well visibly skipped during the serum-pipetting step (Event 1) or the anti-IgM-pipetting step (Event 4), stays colorless; however every serum+anti-IgM well turns the same shade of blue with no visible gradation, so no additional positive/negative intensity distinction is encoded beyond plain presence or absence of color.
level_4: Everything from score 3 holds, including the control well (skipped during serum or anti-IgM pipetting) staying colorless, and blue in the correct wells persists once stop solution is added, but one non-core scientific flaw remains: for example the blue fades again right after stop solution is added instead of holding steady, or the control well shows a faint blue tint caused by motion blur rather than real color development.
level_5: Blue develops only in wells that received serum, anti-IgM-HRP, and TMB substrate, with onset strictly after substrate is added; the control well (skipped during serum or anti-IgM pipetting) stays fully colorless for the entire video; once stop solution is added, the blue in the correct wells holds at a steady intensity with no further darkening, fading, or bleeding into neighboring wells; the causal chain (serum IgM binding the coated antigen, anti-IgM-HRP binding that IgM, and TMB being oxidized only where the enzyme is present) is visible as an unbroken sequence tied to the correct wells alone.

Spatiotemporal Consistency rubric:
level_1: The 96-well plate vanishes from view and reappears in a different spot between shots; the pipette skips instantly from the serum vial to a well with no visible travel between them; liquid inside a well jumps from empty to completely full within a single frame instead of visibly rising as it is dispensed; or the number or layout of wells changes partway through the video.
level_2: At least one major break occurs: the plate suddenly rotates by a large angle with no panning camera motion to account for it; wash fluid disappears mid-pour without draining away; the pipette plunger presses down or releases in sudden jumps rather than a smooth motion; or the room lighting flips abruptly between bright and dim with no visible light-source change to justify it.
level_3: The plate stays centered in frame with the serum, anti-IgM, and substrate vials in fixed positions around it; the pipette travels along one continuous, visible path between vial and well each time; liquid rises gradually and evenly as each reagent is dispensed; and the pipette's cast shadow on the bench stays visible without flickering on and off, but one small inconsistency remains, such as the shadow briefly detaching from the tip for a single frame or a well border flickering for an instant.
level_4: Everything from score 3 holds, and additionally every motion (pipetting, the plate's stillness during each incubation pause, the wash-bottle rinsing) looks mechanically plausible with no floating or sudden reversals, none of the objects change size or warp, and the pipette's shadow on the bench shifts continuously in step with the tip's own motion rather than jumping from one position to another, but one small non-core artifact remains, such as a single frame where the pipette tip briefly appears doubled, or a faint, barely perceptible tremor in the plate during an incubation pause that does not otherwise disrupt the stillness.
level_5: No spatiotemporal break occurs anywhere: the plate stays rigid and flat in view the entire time, the pipette moves along one continuous arc into each well with a natural slowing on contact, liquid enters each well with a visibly rising meniscus rather than snapping to its final level, the pipette's own shadow on the bench and plate tracks its tip continuously from the first pipetting action to the last without ever detaching or jumping, and there is no frame dropping, no doubled objects, no warping, and no moment where the pipette appears to pass through the plate or bench surface.