diff --git a/.changeset/checkbox-ref-callback.md b/.changeset/checkbox-ref-callback.md
new file mode 100644
index 00000000000..f17830c9665
--- /dev/null
+++ b/.changeset/checkbox-ref-callback.md
@@ -0,0 +1,5 @@
+---
+"@primer/react": patch
+---
+
+Checkbox: replace useLayoutEffect with ref callback for setting indeterminate state
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-colorblind-linux.png
index df8374a8184..17b5c878406 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-colorblind-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-colorblind-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-dimmed-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-dimmed-linux.png
index 6c9dcaaed7a..3c80b6af83b 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-dimmed-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-dimmed-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-high-contrast-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-high-contrast-linux.png
index b5be891d721..29bd4ab2aab 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-high-contrast-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-high-contrast-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-tritanopia-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-tritanopia-linux.png
index df8374a8184..17b5c878406 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-tritanopia-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Align-Right-dark-tritanopia-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-colorblind-linux.png
index 40cab8794e3..ee825e631c1 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-colorblind-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-colorblind-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-dimmed-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-dimmed-linux.png
index f428aa05956..8401c455c68 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-dimmed-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-dimmed-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-high-contrast-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-high-contrast-linux.png
index de1cf9e8e55..1b12ea020fb 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-high-contrast-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-high-contrast-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-linux.png
index 40cab8794e3..ee825e631c1 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-tritanopia-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-tritanopia-linux.png
index 40cab8794e3..ee825e631c1 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-tritanopia-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-dark-tritanopia-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-colorblind-linux.png
index 62d614fd193..252456e65ab 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-colorblind-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-colorblind-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-high-contrast-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-high-contrast-linux.png
index 62d614fd193..252456e65ab 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-high-contrast-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-high-contrast-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-linux.png
index 62d614fd193..252456e65ab 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-tritanopia-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-tritanopia-linux.png
index 62d614fd193..252456e65ab 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-tritanopia-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-Responsive-light-tritanopia-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-colorblind-linux.png
index 8470b80931a..582331a0dcd 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-colorblind-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-colorblind-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-dimmed-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-dimmed-linux.png
index 8e6b838735f..c6f3ce0c480 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-dimmed-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-dimmed-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-high-contrast-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-high-contrast-linux.png
index d3912f87d36..74287649a2b 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-high-contrast-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-high-contrast-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-linux.png
index 8470b80931a..582331a0dcd 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-tritanopia-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-dark-tritanopia-linux.png
index 8470b80931a..582331a0dcd 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-colorblind-linux.png
index d530e1ba368..ae53e0d1016 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-high-contrast-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-high-contrast-linux.png
index d530e1ba368..ae53e0d1016 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-linux.png
index d530e1ba368..ae53e0d1016 100644
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index d530e1ba368..ae53e0d1016 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-tritanopia-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Children-light-tritanopia-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-dark-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-dark-colorblind-linux.png
index daf4e252805..b1275421472 100644
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index b59c6974cb6..f797dcbf4d6 100644
Binary files a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-dark-dimmed-linux.png and b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-dark-dimmed-linux.png differ
diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-dark-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-dark-linux.png
index daf4e252805..b1275421472 100644
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index 96883e0c7e4..a88cecef541 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-light-high-contrast-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-Responsive-light-high-contrast-linux.png
index 96883e0c7e4..50b49617076 100644
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index 96883e0c7e4..50b49617076 100644
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index 96883e0c7e4..50b49617076 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-dark-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-dark-colorblind-linux.png
index 40cab8794e3..ee825e631c1 100644
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index f428aa05956..8401c455c68 100644
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index de1cf9e8e55..1b12ea020fb 100644
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index 87b56495dad..ee825e631c1 100644
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index 40cab8794e3..ee825e631c1 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-light-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Custom-Size-On-Parent-light-colorblind-linux.png
index 62d614fd193..252456e65ab 100644
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index 62d614fd193..252456e65ab 100644
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index 62d614fd193..d2a34618dab 100644
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index 62d614fd193..252456e65ab 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Disable-Expand-On-Hover-dark-colorblind-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-Disable-Expand-On-Hover-dark-colorblind-linux.png
index 8470b80931a..2dcacae03bc 100644
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index 8470b80931a..582331a0dcd 100644
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index 2b50fdfb984..5b9cf492ac0 100644
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index 712578f4db4..58fa681ca71 100644
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diff --git a/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-With-Square-Avatars-light-linux.png b/.playwright/snapshots/components/AvatarStack.test.ts-snapshots/AvatarStack-With-Square-Avatars-light-linux.png
index 712578f4db4..e1dd86d01a3 100644
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index 712578f4db4..e1dd86d01a3 100644
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diff --git a/packages/react/src/Checkbox/Checkbox.tsx b/packages/react/src/Checkbox/Checkbox.tsx
index c992f840ebb..3e6ff107e9a 100644
--- a/packages/react/src/Checkbox/Checkbox.tsx
+++ b/packages/react/src/Checkbox/Checkbox.tsx
@@ -1,7 +1,6 @@
 import {clsx} from 'clsx'
-import {useProvidedRefOrCreate} from '../hooks'
-import React, {useContext, useEffect, type ChangeEventHandler, type InputHTMLAttributes, type ReactElement} from 'react'
-import useLayoutEffect from '../utils/useIsomorphicLayoutEffect'
+import {useMergedRefs} from '../hooks'
+import React, {useContext, type ChangeEventHandler, type InputHTMLAttributes, type ReactElement} from 'react'
 import type {FormValidationStatus} from '../utils/types/FormValidationStatus'
 import {CheckboxGroupContext} from '../CheckboxGroup/CheckboxGroupContext'
 import classes from './Checkbox.module.css'
@@ -45,46 +44,48 @@ const Checkbox = React.forwardRef<HTMLInputElement, CheckboxProps>(
     ref,
     // eslint-disable-next-line @typescript-eslint/no-explicit-any
   ): ReactElement<any> => {
-    const checkboxRef = useProvidedRefOrCreate(ref as React.RefObject<HTMLInputElement>)
+    const setIndeterminate = React.useCallback(
+      (node: HTMLInputElement | null) => {
+        if (node) {
+          node.indeterminate = indeterminate || false
+        }
+      },
+      // `checked` is intentionally included even though it is not used inside
+      // the callback: browsers silently clear the `indeterminate` property
+      // whenever the checked state changes, so the callback must re-run to
+      // restore it. The exhaustive-deps rule flags this as an unnecessary
+      // dependency (because `checked` isn't referenced in the body), hence the
+      // suppression below.
+      // eslint-disable-next-line react-hooks/exhaustive-deps
+      [indeterminate, checked],
+    )
+    const mergedRef = useMergedRefs(ref, setIndeterminate)
+
     const checkboxGroupContext = useContext(CheckboxGroupContext)
     const handleOnChange: ChangeEventHandler<HTMLInputElement> = e => {
       checkboxGroupContext.onChange && checkboxGroupContext.onChange(e)
       onChange && onChange(e)
     }
+
+    // aria-checked must reflect three states: mixed (indeterminate), true, or false.
+    const ariaChecked = indeterminate ? ('mixed' as const) : checked ? ('true' as const) : ('false' as const)
+
     const inputProps = {
       type: 'checkbox',
       disabled,
-      ref: checkboxRef,
+      ref: mergedRef,
       checked: indeterminate ? false : checked,
       defaultChecked,
       required,
       ['aria-required']: required ? ('true' as const) : ('false' as const),
       ['aria-invalid']: validationStatus === 'error' ? ('true' as const) : ('false' as const),
+      ['aria-checked']: ariaChecked,
       onChange: handleOnChange,
       value,
       name: value,
       ...rest,
     }
 
-    useLayoutEffect(() => {
-      if (checkboxRef.current) {
-        checkboxRef.current.indeterminate = indeterminate || false
-      }
-    }, [indeterminate, checked, checkboxRef])
-
-    useEffect(() => {
-      const {current: checkbox} = checkboxRef
-      if (!checkbox) {
-        return
-      }
-
-      if (indeterminate) {
-        checkbox.setAttribute('aria-checked', 'mixed')
-      } else {
-        checkbox.setAttribute('aria-checked', checkbox.checked ? 'true' : 'false')
-      }
-    })
-    // @ts-expect-error inputProp needs a non nullable ref
     return <input {...inputProps} className={clsx(className, sharedClasses.Input, classes.Checkbox)} />
   },
 )