Domain: Healthcare — Basic Immunology
Target concept: Inactivation Vaccine Antigenicity Test (Immunofluorescence)

Video generation prompt: Place the antigen-coated slides on a slide rack, add the specific primary antibody, wash, then add the fluorescently labeled secondary antibody, wash again, and observe the slides under a fluorescence microscope. Clearly demonstrate the step-by-step procedures for handling the slides and pipettes, as well as the use of the microscope.

Evaluation guide:
(i) Target concept: Immunofluorescence detection of antigenic integrity via sequential antibody binding.
Minimum mechanism: Antigen must be stably coated on solid phase; primary antibody must be monospecific; secondary antibody must carry a fluorophore and recognize primary antibody species; excitation light must match fluorophore absorption; emitted fluorescence must originate only from bound complexes.
(ii)Storyline: [Fixed overhead + slight frontal angle]. (1) Antigen-coated slide placed onto rack (visible coating texture); (2) Pipette draws primary antibody from labeled vial and deposits it onto slide surface (contact visible); (3) Buffer wash applied across slide (liquid flow visible, no pooling); (4) Pipette draws fluorescent secondary antibody (green-labeled vial) and deposits onto same area; (5) Second wash applied; (6) User positions at fluorescence microscope with UV filter engaged: final frame shows user’s aligned eye and slide’s green emission under excitation (no emission in prior frames). All transitions are continuous, object identities preserved, and causal chain visually anchored: addition → wash → addition → wash → observation → emission.

Manual:
Experimental Procedure
1. Place antigen-coated slide on slide rack.
2. Add specific primary antibody to slide using pipette.
3. Wash slide thoroughly with buffer.
4. Add fluorescently labeled secondary antibody to slide using pipette.
5. Wash slide again. Observe under fluorescence microscope.
Observation of Experimental Phenomena
Bright green-yellow fluorescent signal appears exclusively on antigen-coated regions of the slide after secondary antibody addition and final wash; no signal before secondary addition or on uncoated areas.
Summary of Experimental Principles
Antigen immobilized on slide binds specific primary antibody; fluorescently labeled secondary antibody binds to primary antibody; fluorescence emission under UV excitation confirms antigen-antibody complex formation.

Prompt Grounding rubric:
level_1: No antigen-coated slide, slide rack, pipette, primary antibody container, secondary antibody container, or fluorescence microscope is visible in the scene; or these objects appear but none of the procedural actions occur: no slide placement on the rack, no liquid addition by pipette, no washing motion, and no microscope observation.
level_2: At least one required object (slide, pipette, or microscope) is present, but the core actions are missing or performed in the wrong order, e.g., the secondary antibody is added before the primary antibody, or the microscope is used before any antibody has been applied to the slide; or the correct objects appear on screen but no placement, pipetting, or observation action is ever shown.
level_3: The antigen-coated slide is placed on the slide rack; the primary antibody is added to the slide with a pipette; a washing motion (liquid rinsed across the slide) follows; the secondary antibody is then added with a pipette; and the slide is brought to a fluorescence microscope for observation; all five actions are present, but at least one is imprecise (e.g., the pipette tip is not shown actually touching the slide surface during an addition, or the secondary antibody container is not visually distinguishable from the primary).
level_4: All five actions from the 3-point level occur in the correct order: (1) slide placed on rack, (2) primary antibody added to the slide, (3) washing applied to the slide, (4) fluorescently labeled secondary antibody added to the same slide, (5) user observed at the fluorescence microscope eyepiece, with every object correctly identified (antigen-coated slide, rack, pipette, two visually distinct antibody containers implying different specificities, and a fluorescence microscope with a visible filter turret), and the camera viewpoint stays fixed with consistent spatial relationships between objects throughout.
level_5: All conditions for the 4-point level are met, AND the pipette tip visibly touches the slide surface during each antibody addition rather than dispensing from a height, the wash step shows a continuous stream of liquid moving across the slide rather than a single static droplet, the user's head and posture are fully consistent with looking into the microscope eyepiece (head lowered to eyepiece height, one eye aligned to the lens), AND the primary and secondary antibody containers are clearly visually distinguishable from each other (e.g., different cap colors, vial shapes, or liquid tints) without needing to rely on readable printed text.

Scientific and Causal Correctness rubric:
level_1: No sign of immunofluorescence logic at all: none of the reagents show any color or visual distinction, there is no indication that antigen and antibody are interacting in a specific way, and no fluorescent emission is implied at any point, e.g., every liquid looks like plain colorless water and the primary and secondary antibody containers are visually identical.
level_2: At least one reagent shows some color (e.g., a tinted primary antibody solution), but the primary and secondary antibodies are not visually distinguished from each other, there is no visible consequence of antibody binding, and nothing distinguishes the secondary antibody as fluorescent (no green or yellow glow implied anywhere under the microscope).
level_3: The primary and secondary antibody containers are visually distinct from each other (e.g., different color, tint, or marking suggesting the secondary carries a fluorescent tag), but the slide itself shows no visual evidence that the secondary antibody is actually fluorescent: no green/yellow glow from the slide surface and no excitation light path (e.g., blue/UV illumination) visible at the microscope.
level_4: The secondary antibody container carries an explicit fluorescence cue (e.g., a green tint or a fluorophore name/marking such as FITC or Alexa Fluor 488), the microscope shows a visible excitation light path (e.g., blue-violet illumination or a distinct filter element), and the slide shows a localized bright green-yellow glow only after the secondary antibody has been added and washed, with no such glow visible before the secondary antibody step or on any unlabeled part of the slide.
level_5: All conditions for the 4-point level are met, AND the green-yellow glow appears only on the antigen-coated area of the slide (brighter there than on any bare/uncoated region), the glow is visible only after the final wash step (no lingering diffuse haze right after the secondary antibody is added but before washing), and no green-yellow glow appears from the primary antibody alone or from the wash buffer itself.

Spatiotemporal Consistency rubric:
level_1: Severe spatiotemporal break: the slide, rack, pipette, or microscope vanishes and reappears, or teleports to a different location on screen; the microscope's position or orientation changes discontinuously between frames; room lighting jumps abruptly from bright to dark (or the reverse) between consecutive frames with no transition.
level_2: Moderate rupture: an object moves in a physically impossible way, e.g., the pipette tip passes through the glass slide without any visible contact or deformation, the slide rack rotates more than 90 degrees within a single frame, the wash liquid appears to flow upward against gravity, or the microscope's focus knob visibly turns one way while the image sharpens as though it turned the other way.
level_3: All objects keep consistent relative positions throughout: the slide stays fixed on the rack, the pipette moves continuously toward and away from the slide rather than jumping, the wash liquid follows a smooth downward path across the slide, and the microscope stays in place while the user's head moves smoothly into viewing position, with no vanishing/reappearing objects and no broken or disconnected parts (e.g., the rack's legs stay intact throughout).
level_4: All conditions for the 3-point level are met, AND the motion is physically plausible: the pipette approaches the slide from above at a vertical or shallow angle rather than swiping in sideways, the wash liquid flows downward across the slide surface, the sequence of hand movements matches the order of actions (reaching for a vial, filling the pipette, moving to the slide, then dispensing), and no solid object visibly passes through another (the pipette tip stops right at the slide surface rather than sinking into it).
level_5: All conditions for the 4-point level are met, AND the whole sequence plays out with no jump cuts and no repeated or skipped frames; every motion (e.g., the pipette slowing as it nears the slide, then resuming speed after dispensing) shows smooth acceleration and deceleration; the direction and length of the slide rack's cast shadow stay the same from the start of the clip to the end, the room's overall brightness doesn't visibly flicker, and there is no visible shaking or flicker in the microscope eyepiece alignment as the user looks through it.