diff --git a/extensions/cornerstone/src/utils/hydrationUtils.ts b/extensions/cornerstone/src/utils/hydrationUtils.ts
index 3f6636404..19123ed11 100644
--- a/extensions/cornerstone/src/utils/hydrationUtils.ts
+++ b/extensions/cornerstone/src/utils/hydrationUtils.ts
@@ -76,7 +76,7 @@ function getUpdatedViewportsForSegmentation({
     isHangingProtocolLayout
   );
 
-  if (updatedViewports == null) {
+  if (updatedViewports == null || !isHangingProtocolLayout) {
     return updatedViewports;
   }
 
diff --git a/tests/SEGHydrationFrom3DFourUp.spec.ts b/tests/SEGHydrationFrom3DFourUp.spec.ts
index 78259e584..7c8d5e1d2 100644
--- a/tests/SEGHydrationFrom3DFourUp.spec.ts
+++ b/tests/SEGHydrationFrom3DFourUp.spec.ts
@@ -7,6 +7,7 @@ import {
   visitStudy,
   waitForViewportsRendered,
   waitForViewportRenderCycle,
+  expect,
 } from './utils';
 
 test.beforeEach(async ({ page }) => {
@@ -64,3 +65,54 @@ test.describe('3D four up SEG hydration', async () => {
     });
   });
 });
+
+test.describe('3D four up to 3x2 layout SEG hydration', () => {
+  test.beforeEach(async ({ page }) => {
+    const studyInstanceUID = '1.3.6.1.4.1.14519.5.2.1.256467663913010332776401703474716742458';
+    const mode = 'viewer';
+    await visitStudy(page, studyInstanceUID, mode, 2000);
+  });
+  test('should hydrate SEG only in the target viewport and preserve other viewport orientations', async ({
+    page,
+    DOMOverlayPageObject,
+    leftPanelPageObject,
+    mainToolbarPageObject,
+    viewportPageObject,
+  }) => {
+    await mainToolbarPageObject.layoutSelection.threeDFourUp.click();
+
+    let viewportRenderCycle = waitForViewportRenderCycle(page);
+    // Switch to a manual 3x2 grid layout
+    await mainToolbarPageObject.layoutSelection.grid(3, 2).click();
+    await viewportRenderCycle;
+
+    // Activate the 3rd viewport (index 2) then load the SEG into it
+    await viewportPageObject.getNthLocator(2).click();
+
+    await expect(viewportPageObject.getNthLocator(2)).toHaveAttribute('data-is-active', 'true');
+
+    await leftPanelPageObject.loadSeriesByDescription('SEG');
+
+    await expect(DOMOverlayPageObject.viewport.segmentationHydration.locator).toBeVisible();
+
+    viewportRenderCycle = waitForViewportRenderCycle(page);
+    await DOMOverlayPageObject.viewport.segmentationHydration.yes.click();
+    await viewportRenderCycle;
+
+    await checkForScreenshot({
+      page,
+      locator: viewportPageObject.grid,
+      screenshotPath: screenShotPaths.segHydrationAfterLayoutSwitchTo3By2.afterSEGHydrated,
+    });
+
+    viewportRenderCycle = waitForViewportRenderCycle(page);
+    await mainToolbarPageObject.layoutSelection.threeDFourUp.click();
+    await viewportRenderCycle;
+
+    await checkForScreenshot({
+      page,
+      locator: viewportPageObject.grid,
+      screenshotPath: screenShotPaths.segHydrationAfterLayoutSwitchTo3By2.backTo3DFourUp,
+    });
+  });
+});
diff --git a/tests/pages/MainToolbarPageObject.ts b/tests/pages/MainToolbarPageObject.ts
index 4e3bdf213..434bd53ac 100644
--- a/tests/pages/MainToolbarPageObject.ts
+++ b/tests/pages/MainToolbarPageObject.ts
@@ -30,6 +30,16 @@ export class MainToolbarPageObject {
 
     return {
       ...layoutSelection,
+      grid(cols: number, rows: number) {
+        const button = page.getByTestId(`Layout-${cols - 1}-${rows - 1}`);
+        return {
+          button,
+          async click() {
+            await layoutSelection.click();
+            await button.click();
+          },
+        };
+      },
       get axialPrimary() {
         const button = page.getByTestId('Axial Primary');
         return {
diff --git a/tests/screenshots/chromium/SEGHydrationFrom3DFourUp.spec.ts/afterSEGHydrated.png b/tests/screenshots/chromium/SEGHydrationFrom3DFourUp.spec.ts/afterSEGHydrated.png
new file mode 100644
index 000000000..107995063
Binary files /dev/null and b/tests/screenshots/chromium/SEGHydrationFrom3DFourUp.spec.ts/afterSEGHydrated.png differ
diff --git a/tests/screenshots/chromium/SEGHydrationFrom3DFourUp.spec.ts/backTo3DFourUp.png b/tests/screenshots/chromium/SEGHydrationFrom3DFourUp.spec.ts/backTo3DFourUp.png
new file mode 100644
index 000000000..72463dcbb
Binary files /dev/null and b/tests/screenshots/chromium/SEGHydrationFrom3DFourUp.spec.ts/backTo3DFourUp.png differ
diff --git a/tests/utils/screenShotPaths.ts b/tests/utils/screenShotPaths.ts
index 73cce0481..cee7d87dd 100644
--- a/tests/utils/screenShotPaths.ts
+++ b/tests/utils/screenShotPaths.ts
@@ -151,6 +151,10 @@ const screenShotPaths = {
     threeDFourUpAfterSEG: 'threeDFourUpAfterSEG.png',
     threeDFourUpAfterSegHydrated: 'threeDFourUpAfterSegHydrated.png',
   },
+  segHydrationAfterLayoutSwitchTo3By2: {
+    afterSEGHydrated: 'afterSEGHydrated.png',
+    backTo3DFourUp: 'backTo3DFourUp.png',
+  },
   segNoHydrationThenMPR: {
     segNoHydrationPreMPR: 'segNoHydrationPreMpr.png',
     segNoHydrationPostMPR: 'segNoHydrationPostMpr.png',