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A biological cell is a membrane-bounded basic unit of living organization. In a cell-level experiment it can be an experimental unit, provided that treatment assignment and relevant interactions are handled at that level.

A block design assigns treatments to experimental units partitioned into blocks. Its incidence counts $n_{ij}$ record how often treatment $i$ occurs in block $j$. Balance and connectedness determine which treatment contrasts are estimable and how much adjustment for blocks costs.

A block groups experimental units expected to have similar background responses. Comparing treatments within blocks can reduce the variance of an estimator. Block allocation must be specified before responses are used to evaluate treatment effects.

A completely randomized design assigns treatments directly to available experimental units using complete randomization, without blocking. Fixed group sizes can be enforced. The residual then includes uncontrolled between-unit variation.

Design of experiments chooses experimental units, treatments, replication, blocks in experimental design and randomization so that scientifically useful treatment contrasts can be estimated with meaningful standard errors. Allocation determines which measurements supply independent treatment information.

An experimental unit is a unit to which a treatment is assigned under the experimental allocation. A whole orchard can be an experimental unit even when responses are recorded separately on its trees. Different factors in a split-plot design can have different experimental units.

An observational unit is the object or occasion supplying a recorded response. Several observational units can belong to one experimental unit; counting them as independent treatment replicates creates pseudoreplication.

An orthogonal block design has centered treatment-indicator vectors orthogonal to centered block-indicator vectors on the experimental units. With treatment replication $r_i$, block size $k_j$ and total $n$, this is equivalent to $n_{ij}=r_i{k}_j/n$. It permits additive block adjustment without changing treatment estimates.

An orthogonal block design has treatment contrasts orthogonal to block contrasts after removing the grand mean. This is a statement about vectors on the experimental units, with the usual inner product; the two full spaces are not orthogonal because both contain the constant vector.

For a counting criterion, let $n_{ij}$ count experimental units receiving treatment $i$ in block $j$, let $r_i={\sum }_j{n}_{ij}$, let $k_j={\sum }_i{n}_{ij}$, and let $n={\sum }_i{r}_i$. The inner product of the centered indicators for treatment $i$ and block $j$ is $n_{ij}-r_i{k}_j/n$. ConsequentlyIn particular, equal-sized blocks must contain each treatment in the same proportion. Under an additive block-and-treatment model, adjustment for blocks then does not change the fitted treatment contrasts.

If the biological cells are initially interchangeable, randomly allocate them to five groups of ten for the days. Within each day, independently choose a uniform distribution on a finite set over assignments having two occurrences of each dose level to the ten experimental units. One implementation is a random permutation of a list containing two labels of each level; the duplicate labels give every admissible assignment the same number of underlying permutations.

Randomize the processing order within each day as well, while pairing each preparation with its immediate measurement. Randomize dose within day, not merely the names of the days. This restricted randomization preserves the planned balance while protecting treatment contrasts against systematic order effects. Conceal the dose labels from the assessor where practical.

Use a randomized complete block design, with day as block and each biological cell as an experimental unit. Each day should contain two biological cells at each of the five dose levels. Each level then has ten replicates overall and occurs equally often in every day: the resulting block design is an orthogonal block design.

This spreads any day-to-day changes in preparation, temperature or measurement across all levels instead of confounding dose with day. Treat the zero level as a control with otherwise comparable handling. Keep the treatment-to-measurement delay standardized because the response variable is a diffusion rate measured immediately after preparation.

The experimental unit is a volunteer on a particular afternoon, since that volunteer-session receives a program assignment. There are 36 such experimental units; a volunteer is a repeated block, rather than a unit that receives only one program throughout the experiment.

The observational unit is the individual completed sketch, equivalently the session's single recorded count of correctly represented features. The separate map features contribute to that response variable; they are not independently randomized replicates. Nor are repeated sessions on one volunteer independent subjects.

A completely randomized design uses $2m$ eligible animals, randomly selecting $m$ to receive $P$ and assigning the remainder to $Q$. Use comparable follow-up and assess the same response variable in both groups. Its advantage is simplicity and freedom from previous-treatment carryover effects; its disadvantage is that between-animal variation enters the residual and can make a treatment contrast imprecise.

A randomized complete block design with matched pairs first forms $m$ pairs using pre-treatment characteristics such as initial disease severity, age or breed. Independently choose which animal in each pair receives $P$, with its partner receiving $Q$. The experimental units are animals; pairs are blocks. The average within-pair difference estimates the treatment contrast, and positive within-pair similarity can reduce its variance. Its advantage is control of known heterogeneity; its disadvantage is the need for useful matching, with fewer residual statistical degrees of freedom and little gain if the matching variables are uninformative. Do not construct pairs using outcomes observed after assignment.

A two-period crossover design randomly assigns half the animals to sequence $PQ$ and half to $QP$. Each animal receives both treatments in separate periods, with a scientifically justified interval between them and the same outcome assessment after each period. Animal blocks remove persistent between-animal differences, while the two sequences balance treatment against period. Its advantage is potentially high precision from within-animal comparisons; its disadvantage is vulnerability to carryover effects, changing disease state and irreversible effects. It is suitable only when comparing the treatments in both periods remains meaningful and residual effects of the first treatment are adequately controlled. A two-period crossover design does not by itself disentangle arbitrary treatment-specific carryover effects.

First agree the scientific question and primary response variable. Ask the vet about the disease's course, the proposed mechanism and duration of each treatment, the eligible population, baseline severity, and what outcome and follow-up time would represent worthwhile improvement. Is the disease transmissible, is recovery reversible, and can treating one cow affect another's outcome? These answers determine whether a crossover design is credible, whether the experimental unit should be a cow or a whole herd, and whether individual randomization would leave interference between groups. Agree welfare and rescue arrangements with the vet when deciding which comparisons are feasible; the statistical plan cannot determine these from an unfamiliar disease name.

Second agree feasible allocation and adequate independent replication. Discuss numbers of available cows and herds, variation in the chosen outcome, a scientifically meaningful treatment difference, and the desired statistical power. Use these to plan sample size, rather than choosing a number solely from convenience. Discuss herd, lactation stage and initial severity as potential blocks in experimental design, then specify randomization, comparable management, concealed allocation and blinded outcome assessment where feasible. Repeated milk or health records from the same cow are observational units, not extra independent experimental units; pseudoreplication would give misleading standard errors. Availability of enough independent cows or herds, together with expected variation, governs the precision actually achievable.

Both factors are assigned at orchard level. Therefore the experimental units are the twelve orchards; the trees are observational units within them. The six combinations form a balanced factorial design, replicated twice. The orchard ANOVA stratum has $12-1=11$ statistical degrees of freedom. Spray uses $2-1=1$, pruning uses $3-1=2$, and their interaction term uses $(2-1)(3-1)=2$, leaving six for error.

Dividing each treatment sum of squares in ANOVA by its statistical degrees of freedom and using $1440/6=240$ as the denominator gives all missing entries:

The unrounded F-test statistics are $998/240=4.15833\dots$, $560/240=2.33333\dots$ and $202/240=0.841667\dots$. The within-orchard tree mean square in ANOVA, 180, is not the treatment error denominator: using it would confuse subsampling with independent replication. The tree ANOVA stratum has $12(30-1)=348$ statistical degrees of freedom; $1+11+348=360$ is the uncorrected total, and the corrected total is 359.