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For an original idealized mathematical model, let $n_{{\mathrm{O}}_2}(t)$ be the amount of substance of molecular oxygen consumed since the initial reading and $n_{{\mathrm{CO}}_2}(t)$ the carbon dioxide produced. Suppose a fraction $f$ of that carbon dioxide is removed from the gas, all other gas inventories remain fixed, and temperature $T$ and gas pressure $P$ stay constant. The ideal gas law gives the positive gas contractionwhere $R$ is the molar gas constant and $V$ the gas volume. With complete absorption, $f=1$, the contraction reflects molecular oxygen uptake regardless of the amount of carbon dioxide made. For a uniform pipette bore of area $A$, an inward boundary displacement $x$ gives $D=Ax$. A graduated pipette can instead supply the displaced volume directly.

Let $D_s$ denote sample contraction and $D_b$ the matched blank contraction. Define $D_{\mathrm{corr}}=D_s-D_b$ with both taken relative to their own initial readings. Under the preceding assumptions,The slope of corrected volume against time estimates uptake over the chosen interval. The approximate sign acknowledges that a liquid boundary also responds to hydrostatic and capillary pressure, while the gas can exchange water vapour. A fixed-volume apparatus would instead show a pressure decrease; its ideal conversion is $n_{{\mathrm{O}}_2}\simeq V(P(0)-P(t))/(RT)$ after complete absorption and appropriate blank correction.

The ideal gas law also exposes drift without any biology: for fixed gas inventory, $V\propto T/P$. A one-percent rise in absolute temperature at fixed pressure produces a one-percent expansion. Matching the initial gas spaces makes such common changes easier to subtract. Subtracting unmatched blanks can leave a physical signal that looks biological.

Rolle's theorem is the equal-endpoint case of the mean value theorem. Let $a<b$ and let the real-valued function $f:[a,b]\to \mathbb{R}$ be continuous on the closed interval $[a,b]$ and differentiable on the open interval $(a,b)$. If $f(a)=f(b)$, there exists $c\in (a,b)$ with $f^{\prime }(c)=0$. Geometrically, a tangent line somewhere in the interior is horizontal, like the secant line joining the endpoints. ^[ref]

For the polynomial $f(x)=x(x-2)$ on $[0,2]$, both endpoint values are zero and the derivative is $f^{\prime }(x)=2x-2$. Thus $c=1$, where $f(1)=-1$, and the horizontal tangent line is $y=-1$. The horizontal secant line is $y=0$: equal endpoint values do not force the interior tangent line to pass through those endpoints.

The absolute value function $f(x)=|x|$ on $[-1,1]$ exposes the importance of differentiability. It is continuous and $f(-1)=f(1)=1$, so the secant line has slope zero. However,and the derivative does not exist at zero because the one-sided derivatives disagree. Every existing derivative in the interior is nonzero. The corner fails a hypothesis of both Rolle's theorem and the mean value theorem, so neither conclusion is guaranteed.

For a short proof, the extreme value theorem supplies a greatest and a least value on $[a,b]$. If $f$ is a constant function, every interior point works. Otherwise at least one of these values differs from the common endpoint value, so it is attained at an interior extremum. At an interior local maximum or local minimum, differentiability forces the derivative to be zero: the difference quotients from the two sides have opposite weak signs and the same limit.

This turns repeated values into information about derivatives. Two distinct roots of a polynomial force a root of its derivative between them; iterating this idea gives the Rolle root count with multiplicities. More generally, a differentiable function with nonzero derivative throughout a real interval cannot repeat a value there, hence is injective. The beauty is that knowing only two equal values guarantees an exact local feature somewhere between them.

A secant line is a straight line passing through two distinct points of a curve. For a graph $y=f(x)$ at $x=a$ and $x=a+h$, its slope is the difference quotient $(f(a+h)-f(a))/h$. As the second point approaches the first, a limiting secant line can give the tangent line.

Take identical rigid spheres of radius $a$, particle density ${\rho }_p$ and displaced liquid density ${\rho }_f$, at uniform absolute temperature $T$. Let $z$ increase upwards from a reference layer and write $\Delta \rho ={\rho }_p-{\rho }_f>0$. The buoyancy correction gives the effective gravitational massThe actual grain mass is ${\rho }_{p}V$; the displaced liquid subtracts ${\rho }_{f}V$ from the effective gravitational mass. In a dilute, noninteracting particle suspension, the Boltzmann distribution therefore givesHere $n$ is number density and $k_B$ is the Boltzmann constant. Equivalently, the dilute osmotic pressure $\Pi =n{k}_{B}T$ supports the grains above it: $d\Pi /dz=-n{m}_{b}g$. Dividing by $n$ and integrating yields the same exponential function. No dynamic viscosity appears in this equilibrium argument. If ${\rho }_p<{\rho }_f$, the population instead increases upwards; at equal densities this gravitational model predicts uniform $n$.

A microscope counts a finite optical slab, not an infinitesimal plane. For equal viewed area $A$ and slab thickness $\delta$, an ideal uniform detection window centred at $z$ givesThus equal optical volumes preserve the exponential function count ratio exactly, even for finite $\delta$, provided the windows lie wholly inside the accessible chamber and have the same detection response. Unequal depth response or overlapping a boundary can corrupt this inference. Repeated observations estimate the mean count; closely spaced visits need not be independent because grains remain in view.

For an original chosen model, set $a=0.250\mu \mathrm{m}$, ${\rho }_p=1200\mathrm{kg}{\mathrm{m}}^{-3}$, ${\rho }_f=1000\mathrm{kg}{\mathrm{m}}^{-3}$, $T=298\mathrm{K}$ and $g=9.81\mathrm{m}{\mathrm{s}}^{-2}$. These are illustrative inputs, not Jean Baptiste Perrin's measurements. Using $k_B=1.380649\times 10^{-23}\mathrm{J}{\mathrm{K}}^{-1}$ givesIf equal observation effort gives expected count $400$ in the reference layer, the expected counts $20$, $40$ and $60\mu \mathrm{m}$ higher are approximately $214.3$, $114.8$ and $61.49$. Fractions describe expected values over observations, not fractional grains. Using the actual grain mass instead of its buoyancy-corrected value would make the inferred Avogadro constant six times too small for these chosen densities.

The Seebeck coefficient $S(T)$ quantifies the thermoelectric voltage response per small temperature difference at zero electric current. Use $S=-dV/dT$ along a homogeneous material, with voltage understood through electrochemical potential. Its units are volts per kelvin, commonly $\mu V/K$. The word thermopower is also used, despite this being a voltage response rather than power; some authors use that word for $|S|$, so check their convention. ^[ref].

For the A-minus-B terminal order in the Seebeck effect experiment,Thus a small temperature difference around $T_0$ gives $V_{AB}\approx [S_A(T_0)-S_B(T_0)]\Delta T$. The finite-interval ratio $V_{AB}/\Delta T$ is an average of the coefficient difference, not necessarily its value at either endpoint. Ordinary two-material measurements determine a relative Seebeck coefficient: to recover $S_A$ from the measured difference, add a separately known $S_B$. Neither the choice of a zero for voltage nor the use of a familiar reference material makes $S_B$ identically zero.

A concrete research example is Martin and colleagues' 2021 certification experiment. They measured small temperature differences and voltages on six silicon–germanium specimens at 13 base temperatures between $295$ and $900\mathrm{K}$. At each setting they acquired 40 voltage-and-temperature-difference pairs and used linear regression with a free intercept, so a constant voltage offset did not become part of the slope. Platinum voltage leads required a reference-material correction; their terminal order gave $S_{\mathrm{measured}}=S_{\mathrm{Pt}}-S_{\mathrm{specimen}}$, hence $S_{\mathrm{specimen}}=S_{\mathrm{Pt}}-S_{\mathrm{measured}}$. This is an actual measurement protocol, unlike the toy plot above. ^[ref].

Microscopically, a temperature gradient changes charge carrier transport until an opposing electrical response balances it at zero net electric current. In the usual simple semiconductor picture, electrons and electron holes give opposite Seebeck coefficient signs, and their contributions are weighted by their electrical conductivities. The Seebeck coefficient is a transport property: density of states alone does not determine it, since carrier motion and scattering also matter. Its usefulness extends beyond temperature sensing to diagnosing which carriers transport electric charge. ^[ref].

The Seebeck effect is the generation of a thermoelectric electromotive force by a temperature difference. In a thermocouple, two different homogeneous electrical conductors A and B connect a measuring junction at $T_h$ to two reference connections at a common $T_c$. With negligible electric current, the measured terminal voltage is determined by the difference of their Seebeck coefficients. A hot junction alone does not specify the reading: the reference temperature, the second material, and the terminal order matter.

For a motivating experiment, join one end of A to one end of B, insulating the wires elsewhere. Connect their free ends to identical meter leads at reference temperature $T_c$, for example in an ice-water heat bath. A meter with input electrical resistance much larger than the thermocouple's electrical resistance approximates an open circuit, meaning $I\approx 0$. Keep the measuring junction in a separately controlled heat bath at $T_h$ and wait for stable junction temperatures. Both reference connections must share $T_c$; identical leads with identical endpoint temperatures have cancelling thermoelectric contributions, allowing the meter to be elsewhere. ^[ref].

Define the reading as $V_{AB}=V_A-V_B$, with the positive input on the A reference terminal and the negative input on B. Use the Seebeck coefficient convention in which the voltage change along a homogeneous wire at zero electric current is $dV=-S(T)dT$. Following the path from A at $T_c$ to the common junction at $T_h$, then back along B to $T_c$, givesThis formula assumes homogeneous materials whose Seebeck coefficients depend only on the local temperature, and no additional uncompensated lead contributions. Here voltage is the electrical transport voltage associated with electrochemical potential, rather than a direct measurement of an isolated equilibrium contact electric potential.

Start with $T_h=T_c$, then increase $T_h$ while holding $T_c$ fixed and record $V_{AB}$ at each stable setting. In the ideal model the equal-temperature reading is zero. If $S_A-S_B>0$ throughout the interval, A reads positive when $T_h>T_c$. Exchanging meter inputs reverses the reading; exchanging the two junction temperatures reverses the oriented integral. Repeating the run with identical homogeneous materials gives zero because $S_A=S_B$. These controls test the stated model; a nonzero experimental baseline should be measured rather than silently removed by assuming it is zero.

For an explicitly toy linear example, stipulate constant Seebeck coefficients $S_A=25\mu V/K$ and $S_B=-15\mu V/K$. Choose reference temperature $T_c=20{}^{\circ }\mathrm{C}$ and measuring-junction temperature $T_h=70{}^{\circ }\mathrm{C}$. A temperature difference has the same numerical size in kelvins and on the degree Celsius scale, soSwapping the inputs gives $-2.00\mathrm{mV}$. Bringing both junction regions to the same temperature gives zero. With temperature-dependent Seebeck coefficients, shifting both junction temperatures equally while keeping their difference fixed can change the reading. No actual material pair is claimed to have the stipulated coefficients over this interval.

Real thermocouples need a calibrated voltage-versus-temperature relation: the constant-Seebeck coefficient approximation is useful only where the coefficient difference changes little. Wire inhomogeneity can make the result depend on where the temperature gradient lies, and reference connections at unequal or unknown temperatures introduce errors. An electronic reference correction uses a measured reference temperature in place of an ice bath. ^[ref]. Burns and colleagues provide the standardized reference functions for common letter-designated thermocouple types; the toy line above is not a replacement for them. ^[ref].

The historical experiment used a different detector. In his 1821–1823 investigations, Seebeck found that a closed electrical circuit of two dissimilar materials with junctions at different temperatures deflected a compass needle. The modern explanation is that thermoelectric electromotive force drives an electric current, whose magnetic field deflects the needle. The open-circuit experiment above detects voltage while drawing negligible electric current. The striking connection is that a thermal imbalance can become an electrical signal and then a magnetic response. ^[ref].

A semi-log plot uses a logarithmic scale on one axis and a linear scale on the other. Which axis is logarithmic matters. Plotting $x$ against $\log y$ makes positive exponential functions appear as straight lines; plotting $\log x$ against $y$ instead makes an affine function of a logarithm appear straight. The plotting operations change positions while retaining the original tick values.^[ref]

Take the original dimensionless mathematical model $y(t)=3e^{0.4t}$ for $0\le t\le 8$. It has exponential growth, with $y(0)=3$, $y(4)\approx 14.8591$, and $y(8)\approx 73.5976$. Taking a base-ten logarithm givesThe slope in these transformed positions is $0.4/\ln 10\approx 0.173718$, although the original relative growth rate $y^{\prime }/y$ is $0.4$. Equal steps of two in $t$ multiply $y$ by $e^{0.8}\approx 2.22554$. The corresponding ordinary gains increase, so the left plot bends while the right plot is straight.

One practical use is checking a proposed exponential distribution for failure times. For rate $\lambda >0$ and $t\ge 0$, its cumulative distribution function is $F(t)=1-e^{-\lambda t}$, so the reciprocal survival function $y=1/[1-F(t)]$ satisfies ${\log }_{10}y=\lambda t/\ln 10$. Estimated probabilities can therefore be placed on a logarithmic vertical axis and compared with a straight line. Agreement supports compatibility with the chosen statistical model; a roughly straight finite sample does not establish a unique physical mechanism.^[ref]

In Cartesian coordinates, the slope of a nonvertical straight line is its vertical change divided by its horizontal change. The graph $y=mx+c$ has slope $m$; a vertical straight line has no finite slope. For a tangent line to a differentiable function, the slope is the derivative.

For a nonzero real constant $a$, the tangent line to $y=e^{ax}$ at $x_{\ast }$ passes through the origin precisely when $e^{a{x}_{\ast }}-x_{\ast }a{e}^{a{x}_{\ast }}=0$. Thus $x_{\ast }=1/a$, $y_{\ast }=e$, and its slope is $k=ae$. This gives the unique such tangent line. If $a=0$, the graph is the horizontal line $y=1$ and no tangent line through the origin exists. The equality between the derivative and the slope from the origin, $f^{\prime }(x_{\ast })=f(x_{\ast })/x_{\ast }$, is useful for other differentiable functions too.

A thermocouple reference function gives the nominal thermoelectric electromotive force versus temperature for a specified material pair and reference temperature, conventionally $0{}^{\circ }\mathrm{C}$. Real Seebeck coefficients vary with temperature, sois generally nonconstant. A lookup calibration can use interpolation between tabulated values; a fitted calibration can use polynomials over stated intervals. Use the correct material type, units, reference and validity interval in both the forward and inverse conversions. The FP-TC-120 manual specifically requires the channel's thermocouple type to be configured and describes its use of NIST-175 reference characteristics.^[ref] A nominal reference curve is not evidence that an aged or inhomogeneous wire still follows it.

A local error estimate explains why the reference thermometer matters. If $\delta V$ is an additive voltage error and $\delta T_c$ is the error in the supplied reference temperature, differentiation of the compensation equation gives, for $F^{\prime }(T_h)\ne 0$,A small slope $F^{\prime }(T_h)$ magnifies voltage errors. In the stipulated $40\mu V/K$ model, a $40\mu \mathrm{V}$ offset gives a $1\mathrm{K}$ inferred-temperature error, and a $1\mathrm{K}$ reference error gives a $1\mathrm{K}$ inferred-temperature error. These are sensitivities of the chosen model, not universal performance figures for real sensors.

A trypsin colorimetric assay follows enzyme activity through absorbance at $405\mathrm{nm}$. Trypsin cleaves an amide bond in the synthetic arginine derivative BAPNA, releasing p-Nitroaniline.^[ref]

The inspected Sigma procedure prepares fresh trypsin in cold $1\mathrm{mM}$ hydrochloric acid and fully dissolves $1mg/ml$ BAPNA. Its buffer solution stock contains $200\mathrm{mM}$ triethanolamine and $20\mathrm{mM}$ calcium chloride, at pH $7.8$. Three $3.00\mathrm{ml}$ cuvettes compare trypsin, trypsin plus the enzyme inhibitor aprotinin, and a blank replacing trypsin with its acid solvent. Sodium chloride solution replaces aprotinin in the uninhibited control. A thermostatted instrument for spectrophotometry holds $25{}^{\circ }\mathrm{C}$ and a $1\mathrm{cm}$ light path. After equilibration, adding BAPNA starts approximately five minutes of absorbance recording. The uninhibited sample checks activity; the blank checks background; the inhibited sample tests suppression.^[ref]

Let $s_U$, $s_I$ and $s_B$ denote the measured absorbance slopes per minute over the same linear time interval. Blank subtraction gives $s_U-s_B$ and $s_I-s_B$. If p-Nitroaniline accounts for the changing absorbance, the Beer-Lambert law givesHere $c$ is product molar concentration and $l$ is the geometric light path through the chemical solution. Sigma uses $\epsilon =9.96\mathrm{m}{\mathrm{M}}^{-1}\mathrm{c}{\mathrm{m}}^{-1}$.^[ref]

Choose original illustrative slopes $s_U=0.1036$, $s_I=0.0538$ and $s_B=0.0040\mathrm{mi}{\mathrm{n}}^{-1}$. The corrected uninhibited slope is $0.0996\mathrm{mi}{\mathrm{n}}^{-1}$, giving $dc/dt=0.0100\mathrm{mM}\mathrm{mi}{\mathrm{n}}^{-1}=10.0\mu \mathrm{M}\mathrm{mi}{\mathrm{n}}^{-1}$ at $l=1\mathrm{cm}$. In $3.00\mathrm{ml}$, the product amount of substance increases at $0.0300\mu \mathrm{mol}\mathrm{mi}{\mathrm{n}}^{-1}$. The corrected inhibited slope is $0.0498\mathrm{mi}{\mathrm{n}}^{-1}$, half as large. The relative suppression is thereforeThe background cancels in the numerator, but remains essential in the denominator. Using uncorrected slopes would instead give about $48.07\%$. Differentiating $A_{405}=\epsilon lc$ with fixed $\epsilon$ and $l$ explains the rate conversion; dimensional analysis checks it independently, since $\mathrm{mi}{\mathrm{n}}^{-1}/(\mathrm{m}{\mathrm{M}}^{-1}\mathrm{c}{\mathrm{m}}^{-1}\mathrm{cm})=\mathrm{mM}\mathrm{mi}{\mathrm{n}}^{-1}$.

The inference is a reaction rate on BAPNA under the stated conditions, rather than a direct count of trypsin molecules or proof of every cut in a protein. Cloudiness, other absorbing solutes, or different product protonation can compromise the Beer-Lambert law interpretation. A matched blank and a linear interval are therefore part of the measurement's meaning. A coloured enzyme inhibitor may require its own matched blank; a changed substrate or pH requires a fresh calibration. The constructed constant slopes above describe only that interval, not indefinite product accumulation.

An elastic beam above a smooth obstacle cannot penetrate it, and a nonadhesive obstacle can only push. The displayed constraints express this when $R$ is an upward distributed normal contact force; concentrated point forces must also be nonnegative. In the small-slope Euler-Bernoulli beam equation,Integrating across a contact point with no applied couple gives jump conditions $[w]=[w^{\prime }]=[B{w}^{\prime \prime }]=0$ and $[B{w}^{\prime \prime \prime }]=R_i$, provided the tension is continuous. For a flat obstacle, no distributed load and zero tension, an open contact interval has $R=0$; its entire normal contact force can therefore be concentrated at its endpoints.